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| − | Penn iGEM 2015 Daily Digest
| + | <h3 style= "text-align: center;">Week 1</h3> |
| − | </p> | + | |
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| − | <a href="https://docs.google.com/document/d/1HvOEUNTXplBFVLyiLFqUM917iPTD_HMbZdsruq0erJE/edit#heading=h.sxfktjtcmvnc">Week 2 (6/8 - 6/12)</a>
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| − | <a href="https://docs.google.com/document/d/1HvOEUNTXplBFVLyiLFqUM917iPTD_HMbZdsruq0erJE/edit#heading=h.bhn2oj1e6cnr">Week 3 (6/15 - 6/19)</a>
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| − | <a href="https://docs.google.com/document/d/1HvOEUNTXplBFVLyiLFqUM917iPTD_HMbZdsruq0erJE/edit#heading=h.xgegbjwvfyf">Week 4 (6/22 - 6/26)</a>
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| − | <a href="https://docs.google.com/document/d/1HvOEUNTXplBFVLyiLFqUM917iPTD_HMbZdsruq0erJE/edit#heading=h.3ael4skbu2k4">Week 5 (6/29 - 7/3)</a>
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| − | <a href="https://docs.google.com/document/d/1HvOEUNTXplBFVLyiLFqUM917iPTD_HMbZdsruq0erJE/edit#heading=h.gvbqdzb1kqf6">Week 6 (7/6 - 7/10)</a>
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| − | <a href="https://docs.google.com/document/d/1HvOEUNTXplBFVLyiLFqUM917iPTD_HMbZdsruq0erJE/edit#heading=h.ebadnxicnqsi">Week 7 (7/13 - 7/17)</a>
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| − | <a href="https://docs.google.com/document/d/1HvOEUNTXplBFVLyiLFqUM917iPTD_HMbZdsruq0erJE/edit#heading=h.qjnk87xe52sy">Week 8 (7/20 - 7/24)</a>
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| − | <a href="https://docs.google.com/document/d/1HvOEUNTXplBFVLyiLFqUM917iPTD_HMbZdsruq0erJE/edit#heading=h.9ytnncddh38t">Week 9 (7/27 - 7/31)</a>
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| − | <a href="https://docs.google.com/document/d/1HvOEUNTXplBFVLyiLFqUM917iPTD_HMbZdsruq0erJE/edit#heading=h.v1nevkzea28q">Week 10 (8/3 - 8/7)</a>
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| − | <a href="https://docs.google.com/document/d/1HvOEUNTXplBFVLyiLFqUM917iPTD_HMbZdsruq0erJE/edit#heading=h.x44heahdq0mm">-Skip Week 11-</a>
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| − | <a href="https://docs.google.com/document/d/1HvOEUNTXplBFVLyiLFqUM917iPTD_HMbZdsruq0erJE/edit#heading=h.nm5utcm00u80">Week 12 (8/17 - 8/21)</a>
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| − | <br/>
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| − | <h1 dir="ltr">
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| − | Week 2 (6/8 - 6/12)
| + | |
| − | </h1>
| + | |
| − | <p dir="ltr">
| + | |
| − | 6/8/2015
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for the day:
| + | |
| − | </p> | + | |
| | <ol> | | <ol> |
| − | <li dir="ltr">
| + | <li>Molecolar Biology Training Workshop</li> |
| − | <p dir="ltr">
| + | <li>Practiced the basics of molecular cloning</li> |
| − | Run a gel after digestion of lacZ and pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr"> | + | |
| − | <p dir="ltr">
| + | |
| − | Gel extraction & digest cleanup
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Ligation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transformation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| | </ol> | | </ol> |
| − | <br/> | + | <h3 style= "text-align: center;">Week 2</h3> |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p> | + | |
| | <ol> | | <ol> |
| − | <li dir="ltr"> | + | <li>Idea Brainstorming and Generation</li> |
| − | <p dir="ltr">
| + | <li>Compiled a preliminary list of potential ideas</li> |
| − | Ran a gel after digestion
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Performed gel extraction and digestion cleanup
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr"> | + | |
| − | <p dir="ltr">
| + | |
| − | Measured concentrations of pDawn and lacZ with nanodrop
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| | </ol> | | </ol> |
| − | <br/> | + | <h3 style= "text-align: center;">Week 3</h3> |
| − | <p dir="ltr">
| + | |
| − | Aims for tomorrow:
| + | |
| − | </p> | + | |
| | <ol> | | <ol> |
| − | <li dir="ltr"> | + | <li>Settled on two main ideas: 1. Quorum sensing with antibiotics 2. Heavy metal removal with Magnetotactic bacteria</li> |
| − | <p dir="ltr">
| + | <li>Learned how to use Geneious to design cloning process</li> |
| − | PCR clean-up for new PCR of lacZ
| + | <li>Practiced extracting biobricks and transforming NEB Turbo cells with plasmids</li> |
| − | </p>
| + | |
| − | </li>
| + | <li>Human Practices</li> |
| − | <li dir="ltr"> | + | <li>Visited Biomeme to get the portable qPCR machine</li> |
| − | <p dir="ltr">
| + | |
| − | Digestion of lacZ and pDawn with new Nde1 and BamHI
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr"> | + | |
| − | <p dir="ltr">
| + | |
| − | Digest clean-up
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr"> | + | |
| − | <p dir="ltr">
| + | |
| − | Ligation of lacZ and pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr"> | + | |
| − | <p dir="ltr">
| + | |
| − | Lux box transformation into E. coli
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| | </ol> | | </ol> |
| − | <br/> | + | <h3 style= "text-align: center;">Week 4</h3> |
| − | <p dir="ltr">
| + | |
| − | 6/9/2015
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for today:
| + | |
| − | </p> | + | |
| | <ol> | | <ol> |
| − | <li dir="ltr"> | + | <li>Decided on the project idea: Heavy metal removal with Magnetotactic bacteria |
| − | <p dir="ltr">
| + | <li>Identified primers and promoters in AMB-1 strain |
| − | PCR cleanup for lacZ
| + | <li>Identified AMB-1 transformation vector |
| − | </p>
| + | <li>Extracted smtA biobrick gene and made glycerol stock |
| − | </li> | + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digest lacZ and pDawn
| + | |
| − | </p>
| + | |
| − | </li> | + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digest cleanup
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Ligation of pDawn and lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux box transformation
| + | |
| − | </p>
| + | |
| − | </li> | + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Design one plasmid system with Lux box/lacZ and design two plasmid system, one with Lux box and the other with the lacZ reporter
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| | </ol> | | </ol> |
| − | <br/> | + | <h3 style= "text-align: center;">Week 5</h3> |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <br/> | + | |
| | <ol> | | <ol> |
| − | <li dir="ltr"> | + | <li>Developed three goals to accomplish for the project |
| − | <p dir="ltr">
| + | <li>Determined the constructs to clone into AMB-1 |
| − | PCR cleanup for lacZ
| + | <li>Designed two fast-fail experiments |
| − | </p>
| + | <li> Created a workflow for the construction of plasmid |
| − | </li> | + | |
| − | <li dir="ltr"> | + | <li> Human Practices |
| − | <p dir="ltr">
| + | <li>Planned outreach events at high school summer programs |
| − | Digest of lacZ and pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digest cleanup of lacZ and pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Ligation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux box transformation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Began working on plasmid design
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr"> | + | |
| − | <p dir="ltr">
| + | |
| − | Inoculated pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| | </ol> | | </ol> |
| − | <br/> | + | <h3 style= "text-align: center;">Week 6</h3> |
| − | <p dir="ltr">
| + | |
| − | Aims for tomorrow:
| + | |
| − | </p> | + | |
| | <ol> | | <ol> |
| − | <li dir="ltr"> | + | <li>Ordered chemicals for AMB-1 growth medium |
| − | <p dir="ltr">
| + | <li>Completed primer design |
| − | Potentially design primers for cloning Lux box into pDawn/lacZ
| + | <li>Obtained AMB-1 strain from Dr.Goolian |
| − | </p>
| + | |
| − | </li> | + | |
| − | <li dir="ltr"> | + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Design SapI primers
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Design compatibility system
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pick colonies for BioBrick, inoculate, glycerol stock
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Colony PCR pDawn + lacZ, glycerol stock, miniprep, sequencing
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Induce lux box w/ arabinose
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| | </ol> | | </ol> |
| − | <br/> | + | <h3 style= "text-align: center;">Week 7</h3> |
| − | <p dir="ltr">
| + | |
| − | Questions:
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | In the “empty” region of pDawn (the only space we think a biobrick can be cloned) there’s only
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | one restriction enzyme that cuts once in that region and nowhere else, SapI.
| + | |
| − | </p>
| + | |
| − | <br/> | + | |
| | <ol> | | <ol> |
| − | <li dir="ltr"> | + | <li>Determined the OD600 plate reading conversion formola experimentally |
| − | <p dir="ltr">
| + | <li>Attempted to do E.Coli and AMB-1 cadmium tolerance test |
| − | Are we right in assuming that empty space is the only place we can clone in a biobrick?
| + | <li>Designed all construct on Geneious |
| − | </p>
| + | |
| − | </li> | + | <li>Human Practices |
| − | <li dir="ltr"> | + | <li>Met with Dr.Rizk to discuss presenting to incoming bioengineering freshmen |
| − | <p dir="ltr">
| + | |
| − | Would it be okay if we used only one restriction enzyme for the ligation instead of two?
| + | |
| − | </p>
| + | |
| − | </li> | + | |
| | </ol> | | </ol> |
| − | <br/> | + | <h3 style= "text-align: center;">Week 8</h3> |
| − | <p dir="ltr">
| + | |
| − | 6/10/2015
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for today:
| + | |
| − | </p> | + | |
| | <ol> | | <ol> |
| − | <li dir="ltr"> | + | <li>Ordered DNAs from Genscript and IDT |
| − | <p dir="ltr">
| + | |
| − | Miniprep pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Design SapI primers
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Design Compatibility Systems
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pick Colonies for Biobrick, inoculate, glycerol stock
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Colony PCR, pDawn + LacZ, Glycerol stock, miniprep
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Redigest pDawn (with extreme controls)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Reattempt Ligation of pDawn+LacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Colony PCR of BioBrick Transformation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run gel of colony PCR products
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run gel of newly digested pDawn, Overnight ligation.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| | </ol> | | </ol> |
| − | <br/> | + | <h3 style= "text-align: center;">Week 9</h3> |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p> | + | |
| | <ol> | | <ol> |
| − | <li dir="ltr"> | + | <li>Redid E.Coli Cadmium Tolerance Test in M9 media |
| − | <p dir="ltr">
| + | <li>Learned to do cell count for AMB-1 |
| − | Miniprep pDawn
| + | <li>Attempted to make AMB-1 chemically competent |
| − | </p>
| + | |
| − | </li> | + | <li>Human Practices |
| − | <li dir="ltr">
| + | <li>Presented to high school students at Penn M&T program |
| − | <p dir="ltr">
| + | <li>Planned a synthetic biology preceptorial |
| − | Re-digest of pDawn (with extreme controls)
| + | <li>Contacted Schuylkill Action Network |
| − | </p>
| + | |
| − | </li> | + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Re-attempt Ligation of pDawn+LacZ
| + | |
| − | </p>
| + | |
| − | </li> | + | |
| − | <li dir="ltr"> | + | |
| − | <p dir="ltr">
| + | |
| − | Colony PCR of pDawn + LacZ
| + | |
| − | </p>
| + | |
| − | </li> | + | |
| − | <li dir="ltr"> | + | |
| − | <p dir="ltr">
| + | |
| − | Gel extract of newly digested pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transformed lux box BioBrick, pDawn + lacZ, just pDawn backbone (for miniprep and
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | making our own glycerol stock) into NEB turbo again [had just cut pDawn backbone and just digested lacZ insert transformed as well as controls]
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plated transformed colonies and placed into incubator (will look at results tomorrow)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| | </ol> | | </ol> |
| − | <br/> | + | <h3 style= "text-align: center;">Week 10</h3> |
| − | <p dir="ltr">
| + | |
| − | Aims for tomorrow:
| + | |
| − | </p> | + | |
| | <ol> | | <ol> |
| − | <li dir="ltr"> | + | <li>Completed AMB-1 growth curve under different media |
| − | <p dir="ltr">
| + | <li>Determined cell count and OD600 conversion formola for AMB-1 experimentally |
| − | Run gels of colony PCR from 6/10/15 and see if the lacZ was ligated into pDawn backbone
| + | <li>Completed E.Coli Cadmium Tolerance Test in LB media |
| − | </p>
| + | |
| − | </li>
| + | <li>Human Practices |
| − | <li dir="ltr">
| + | <li> Volunteered at SEA Science Carnival |
| − | <p dir="ltr">
| + | |
| − | Finish designing cloning of luxbox biobrick into pACBB-eGFP (plasmid that is compatible with pDawn)
| + | |
| − | </p>
| + | |
| − | </li> | + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pick colonies for most recent BioBrick Transformation, Inoculate, glycerol stock, miniprep
| + | |
| − | </p>
| + | |
| − | </li> | + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Colony PCR of new pDawn + LacZ transformation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Induce luxbox transformed E. coli with arabinose (growing up transformed E. coli to saturation, back diluting 1:100, growing up to OD600 of 0.3,
| + | |
| − | then inducing with low to high arabinose concentrations)
| + | |
| − | </p>
| + | |
| − | </li> | + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | If transformation results not looking good, we will need to re-digest lacZ insert and pDawn, and re-ligate pDawn into lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| | </ol> | | </ol> |
| − | <br/> | + | <h3 style= "text-align: center;">Week 11</h3> |
| − | <p dir="ltr">
| + | |
| − | Questions:
| + | |
| − | </p> | + | |
| | <ol> | | <ol> |
| − | <li dir="ltr"> | + | <li>Addressed EMSGM growth problem with pH |
| − | <p dir="ltr">
| + | <li>Make new recovery media with adjusted pH for optimal AMB-1 growth |
| − | How exactly does the two-plasmid transformation work? Do we transform the NEB turbo with one and then make it competent again and transform it with
| + | <li>Chemical Transformation with different recovery broths |
| − | the second?
| + | <li>Attempted PCR assembly with synthesized parts |
| − | </p>
| + | |
| − | </li> | + | |
| − | <li dir="ltr"> | + | |
| − | <p dir="ltr">
| + | |
| − | Is there any way we can check for incomplete digestion (because the gel resolution doesn't allow us to see that, since the part we cut out is only
| + | |
| − | a few bases big)?
| + | |
| − | </p>
| + | |
| − | </li> | + | |
| | </ol> | | </ol> |
| − | <br/> | + | <h3 style= "text-align: center;">Week 12</h3> |
| − | <p dir="ltr">
| + | |
| − | 6/11/2015
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for today:
| + | |
| − | </p> | + | |
| | <ol> | | <ol> |
| − | <li dir="ltr"> | + | <li>Used anaerobic chambers to grow plates |
| − | <p dir="ltr">
| + | <li>Troubleshooted lack of magnetism with new iron maleate solution |
| − | Inoculate pDawn
| + | <li>Troubleshooted AMB-1 electroporation experiment with plating at every step |
| − | </p>
| + | <li>Began cloning successfolly assembled PCRed constructs into PYMB essentials |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run gel of colony PCR
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Check plates from yesterday
| + | |
| − | </p>
| + | |
| − | </li> | + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Colony PCR of re-transformed plates (after we miscalculated the ligation of the first plates)
| + | |
| − | </p>
| + | |
| − | </li> | + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digest and ligate pACBB-eGFP and lux box
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up luxbox
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculate pACBB-eGFP
| + | |
| − | </p>
| + | |
| − | </li> | + | |
| | </ol> | | </ol> |
| − | <br/> | + | <h3 style= "text-align: center;">Week 13</h3> |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p> | + | |
| | <ol> | | <ol> |
| − | <li dir="ltr"> | + | <li>Finished cloning flurescent protein with smtA and mCherry into PYMB essentials |
| − | <p dir="ltr">
| + | <li>Transform AMB-1 with this construct to test for successfol transformation with fluorescence test |
| − | Inoculated pDawn
| + | |
| − | </p>
| + | |
| − | </li> | + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Ran gel of first colony PCR
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Checked plates grown overnight
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Performed second colony PCR of newly transformed plates with pDawn + lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grew up luxbox transformants in LB+chlor (see double check in transformation worked because Chlor diffused in plate)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculated DH5alpha with pACBB-eGFP (in prep for miniprep and glycerol stock tomorrow)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Retransformed lux box biobrick into NEB turbo
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| | </ol> | | </ol> |
| − | <br/> | + | <h3 style= "text-align: center;">Week 14</h3> |
| − | <p dir="ltr">
| + | |
| − | Aims for tomorrow:
| + | |
| − | </p> | + | |
| | <ol> | | <ol> |
| − | <li dir="ltr"> | + | <li>Make new E-MSGM media |
| − | <p dir="ltr">
| + | <li>Re-designed construct |
| − | Miniprep and glycerol stock pACBB-eGFP inoculants
| + | |
| − | </p>
| + | |
| − | </li> | + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Check inoculated colonies from luxbox transformation attempt #2 to see if any grew in LB+Chlor
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Look at transformation results on plates (for attempt #3 of transforming lux box biobrick)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Look at results for colony PCR of pDawn + lacZ (run gel + image)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| | </ol> | | </ol> |
| − | <br/> | + | <h3 style= "text-align: center;">Week 15</h3> |
| − | <p dir="ltr">
| + | |
| − | Questions:
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | None, we might have some after we get the colony PCR results.
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 6/12/2015
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for today:
| + | |
| − | </p> | + | |
| | <ol> | | <ol> |
| − | <li dir="ltr"> | + | <li>Troubleshooted cloning E. coli construct for cadmium tolerance |
| − | <p dir="ltr">
| + | <li>Sequence verified finished constructs on PYMB |
| − | Miniprep pACBB-eGFP
| + | |
| − | </p>
| + | |
| − | </li> | + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock of pACBB-eGFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Back dilute/ induce lux box→ Back dilute @ 12:57 PM
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Colony PCR lux box→ run on a gel
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock lux box
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock of pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 3 hr digest of pDawn + lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Overnight ligation of pDawn + lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run colony PCR on a gel for pDawn + lacZ ligation product.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| | </ol> | | </ol> |
| − | <br/> | + | <h3 style= "text-align: center;">Week 16</h3> |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p> | + | |
| | <ol> | | <ol> |
| − | <li dir="ltr"> | + | <li>Built the prototype of a portable spectrophotometer for cell recovery experiment |
| − | <p dir="ltr">
| + | <li>Designed primers for Gibson Assembly of final construct |
| − | pCR lacZ
| + | |
| − | </p>
| + | <li>Human Practices |
| − | </li>
| + | <li>Presented to Penn BE 100 lecture to intriduce synthetic biology to freshman bioengineers. |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep pACBB-eGFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock of pACBB-eGFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Colony PCR lux box→ run on a gel
| + | |
| − | </p>
| + | |
| − | </li> | + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock of pDawn
| + | |
| − | </p>
| + | |
| − | </li> | + | |
| − | <li dir="ltr"> | + | |
| − | <p dir="ltr">
| + | |
| − | Run colony PCR on a gel for pDawn + lacZ ligation product.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| | </ol> | | </ol> |
| − | <br/> | + | <h3 style= "text-align: center;">Week 17</h3> |
| − | <p dir="ltr">
| + | |
| − | Aims for Monday:
| + | |
| − | </p> | + | |
| | <ol> | | <ol> |
| − | <li dir="ltr"> | + | <li>Tested AMB-1 aerobic colture and anaerobic colture with magnetometer |
| − | <p dir="ltr">
| + | <li>Troubleshooted and retried cloning E. coli construct |
| − | Induce lux box
| + | <li>PCRed up parts we received for construction of shuttle vector |
| − | </p>
| + | |
| − | </li> | + | |
| − | <li dir="ltr"> | + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock lux box
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 3 hr digest of pDawn + lacZ (didn’t do before because needed more lacZ)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Overnight ligation of pDawn + lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Clone lux box into pACBB-eGFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| | </ol> | | </ol> |
| − | <br/> | + | <h3 style= "text-align: center;">Week 18</h3> |
| − | <p dir="ltr">
| + | |
| − | Questions:
| + | |
| − | </p> | + | |
| | <ol> | | <ol> |
| − | <li dir="ltr"> | + | <li>Tested AMB-1 aerobic colture and anaerobic colture with magnetometer |
| − | <p dir="ltr">
| + | <li>Finished PCRing parts we received from IDT |
| − | Does completing PCR on LacZ as a positive control using the Colony PCR Protocol (a.k.a using Taq instead of Phusion) have a significant effect on
| + | |
| − | the results of the PCR? [If not clear, this question is better explained in person]
| + | |
| − | </p>
| + | |
| − | </li> | + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Should we purchase the enzymes and dntps? How long after the expiration date are the enzymes good for? Should we get rid of enzymes/ dntps that are
| + | |
| − | months/years past their expiration date?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | What happens if you use a buffer for restriction digest that is not the optimal one?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| | </ol> | | </ol> |
| − | <br/> | + | <h3 style= "text-align: center;">Week 19</h3> |
| − | <h1 dir="ltr">
| + | |
| − | Week 3 (6/15 - 6/19)
| + | |
| − | </h1>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | -6/15/2015 SKIPPED-
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | 6/16/2015
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for today:
| + | |
| − | </p>
| + | |
| − | <br/> | + | |
| | <ol> | | <ol> |
| − | <li dir="ltr">
| + | <li> Measured the magnetic strength indicator T2 for various concentration of AMB-1 colture and attempted to develop a trendline |
| − | <p dir="ltr">
| + | <li> Counted folly grown sample of aerobic AMB-1 and made a trendline converting OD to cell concentration |
| − | Innoculate pACBB
| + | <li>Cloned all biobricks into PSB1C3 backbone |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make M9 media
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Perform gel extraction on pDawn, lacZ, pACBB-eGFP, and luxbox from gel we ran yesterday.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Ligate luxbox and pACBB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | LIgate pDawn and lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform luxbox and pACBB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform pDawn and lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Back dilute and induce luxbox
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Measure luminescence of induced luxbox on the hour
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | PCR cleanup on lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 3 hour digest of lacZ
| + | |
| − | </p>
| + | |
| − | </li> | + | |
| | </ol> | | </ol> |
| − | <br/> | + | <h3 style= "text-align: center;">Week 20</h3> |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <br/> | + | |
| | <ol> | | <ol> |
| − | <li dir="ltr"> | + | <li>compiled all data and graphics for wiki |
| − | <p dir="ltr">
| + | |
| − | Made M9
| + | |
| − | </p>
| + | </ol><div id="textbox"> |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Gel extracted pDawn, lacZ, luxbox, and pACBB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Ligated luxbox and pACBB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Ligated pDawn and lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transformed luxbox and pACBB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transformed pDawn and lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Back diluted and induced luxbox
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Measured luminescence of induced luxbox on the hour
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | PCR clean up on lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digested lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Innoculate pACBB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for tomorrow:
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock luxbox and pSB1C3
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep more pACBB-eGFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Look at transformed plates, pick colonies, innoculate
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Colony PCR lacZ from pDawn and luxbox from pACBB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Column purify digested lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Dilute luxbox to OD600 .3, take measurements at 1 hr, 3 hr, 5 hr, etc.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Take 24th hour time point for lux box trial 1 (the one we started today)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Potentially do overnight ligations if the transformations from today don’t work
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep and glycerol stock ligated plasmids pDawn and lacZ and luxbox and pACBB.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 6/17/2015
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock LuxBox + PSB1C3
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep more pACBB-eGFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Look at plates, pick colonies, inoculate at 4pm LuxBox+pACBB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Colony PCR luxbox from pACBB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Column purify digested lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform overnight ligation (& prep plates)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | miniprep and glycerol stock ligated pDawn+LacZ and Luxbox+pACBB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform BL21 with luxbox biobrick.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock of LuxBox+PSB1C3
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep more pACBB-eGFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Design primer for lacZ RE site extensions
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Design luxbox colony PCR primers
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Design luxbox sequencing primers
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Design lacZ sequencing primers
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Look at plates (ligation products lacZ + pDawn and lux + pACBB), pick colonies, innoculate.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plot data of lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculate BL21 w/ lux biobrick (to back dilute on Fri)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Friday:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep + glycerol stock lacZ + pDawn and lux + pACBB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Dilute BL21 w/ lux biobrick to OD600 .3
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Take measurements at 1 hr, 3 hr, 5 hr, etc.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plot data ^
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Questions:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Is N = 2 enough for NEB Turbo for measuring luminescence experiment (as opposed to BL21 which we will do later)? Should we do more trials?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Should we do the interlab study this year? ( <a href="https://2015.igem.org/Tracks/Measurement/Interlab_study">https://2015.igem.org/Tracks/Measurement/Interlab_study</a>)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | We should discuss human practices
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 6/18/2015
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculate BL21 LuxBox + PSB1C3
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Colony PCR of pDawn + LacZ transformation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculate pACBB + LuxBox, pDawn + LacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform pDawn + LacZ from Mike into Turbo Cells
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digest test for pDawn + LacZ; pACBB + Lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol Stock BL21 with LuxBox
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order LacZ Assay Kit
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order pDawn & pACBB insert primers
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order Lux Sequence primers.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make more arabinose, autoclave flasks, prep for BL21 vs. Turbo experiment.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make LB Chlor. plates
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculate BL21 LuxBox + PSB1C3
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculate pACBB + LuxBox, pDawn + LacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform pDawn + LacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order pDawn & pACBB insert primers
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Find LacZ Assay kit we want to order (See links below)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order Lux Sequence primers.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make more arabinose, autoclave flasks, prep for BL21 vs. Turbo experiment.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol Stock BL21 + LuxBox
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digest Test of pDawn + LacZ & pACBB + LuxBox
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave LB Agar & pour plates ( Add Chlor)!!
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Back dilute & induce BL21 and NEB Lux strains
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Take luminescence measurements.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 6/19/2015
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol Stock BL21 + LuxBox
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digest Test of pDawn + LacZ & pACBB + LuxBox
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave LB Agar & pour plates
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pick colonies pDawn + lacZ; inoculate
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock pDawn + lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock 6 colonies for BL21 vs turbo experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol Stock BL21 + LuxBox
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digest Test of pDawn + LacZ & pACBB + LuxBox
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave LB Agar & pour plates
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pick colonies pDawn + lacZ; inoculate
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock pDawn + lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock 6 colonies for BL21 vs turbo experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Monday:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Back dilute & induce BL21 and NEB Lux strains
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Take luminescence measurements.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculate 10 ml cultures
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <h1 dir="ltr">
| + | |
| − | Week 4 (6/22 - 6/26)
| + | |
| − | </h1>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Back-dilute to 0.3
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Measure luminescence at 1h, 3h, etc.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Presentation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | lacZ expression as a result of light
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB vs. PSBIC3
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | BL21 vs. turbo in lux exp.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | lux exp. high vs low AHL →
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | questions
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | GANT for summer
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 20 kb from IDT
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | nonhydrolyzable arabinose
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Josh + Danny at 1 PM
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Decide if digest test worked
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol start="6">
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Back-dilute to 0.3
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Measure luminescence at 1h, 3h, etc.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Presentation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | lacZ expression as a result of light
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB vs. PSBIC3
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | BL21 vs. turbo in lux exp.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | lux exp. high vs low AHL →
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | questions
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | GANT for summer
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 20 kb from IDT
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | nonhydrolyzable arabinose
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Josh + Danny at 1 PM
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Decide if digest test worked
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Monday:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze luxbox in BL21 versus NEB Turbo
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform ligated pDawn+lacZ and pACBB+lux, then induce with arabinose
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Explore IDT 20kb options
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Figure out our goals (given our knowledge of Peking)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Work on website design
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Work on human practices
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 6/23/15
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims For Today:
| + | |
| − | </p>
| + | |
| − | <ol start="7">
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze luxbox in BL21 versus NEB Turbo
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform ligated pDawn+lacZ and pACBB+lux, then induce with arabinose
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Explore IDT 20kb options
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Figure out our goals (given our knowledge of Peking)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Work on website design
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Work on human practices
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplished Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyzed luxbox in BL21 versus NEB Turbo data
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transformed pDawn+lacZ and pACBB+lux into BL21
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Looked into other past iGEM team projects involving communication through light
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Started thinking about human practices and figuring out the logistics of sorting past projects by topic/team/year
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims For Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Compare fluorescent output for BL21 at different ODs by taking measurements 1hr, 3hr, 5hr, etc.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Questions/Concerns:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Discuss what the other teams are doing/have done and how are goals should shift accordingly
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 6/24/15
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims For Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Back dilute luxbox cultures to different OD’s, induce, measure OD until 7 hrs
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Decide on 20 kb from IDT
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Listen to IDT webinar @2 pm
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Look @ transformed plates and innoculate
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock ligated pDawn + lacZ and pACBB + lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Look over lacZ kit
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up cultures for both (ours and Mike’s) pDawn lacZ experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock Mike’s pDawn+lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up lux BL21 cultures for arabinose spiking experiments
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order d-fucose
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims For Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn+lacZ experiment (varying blue light intensity)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | back dilute lux BL21 to .3 and induce with arabinose
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Measure lux BL21 and add arabinose every hour
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pick up dNTPs
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 6/25/15
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims For Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Re-do yesterday’s experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pick up dNTPs
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | order more SOC
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | make color chart _ plan prospective experiments
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | IDT → $$
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | plan lacZ + pDawn experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Research previous team projects
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Created a spreadsheet detailing all sender-xreceiver cell relationships
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Rethought lux box experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Set up tomorrow’s pDawn + lacZ experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculated cultures for ^ experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Complete pDawn + lacZ experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pick up dNTPs
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order more SOC
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | IDT → $$
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Karol’s iGEM database project
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 6/26/15
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims For Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | iGEM database (human practices)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | B-Galactosidase assay
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finalize plan for the summer
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | OD + luminescence experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | iGEM database (human practices)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | B-Galactosidase assay
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | OD + luminescence experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Monday:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Co-transforming pDawn + lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Redoing standardized curve for B-Galactosidase assay
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Design exp for different blue light intensity on B- Galactosidase activity experiment.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + pSB1C3 experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Did pACBB + lux ligate properly? (we learned that pDawn + lacZ cloning did not work, but we still have Mike’s which turned yellow when we did
| + | |
| − | the assay yay!)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Decide which parts to order from IDT (but actually though)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Examine more of the different color systems that we have been discussing; talk to advisors about this more; decide which we want to do
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Questions:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | <a href="https://docs.google.com/spreadsheets/d/1Hos0nFdv6kmtn_Q1y98bH6RwkHVTM650sZQLPafrFRg/edit?usp=sharing">
| + | |
| − | https://docs.google.com/spreadsheets/d/1Hos0nFdv6kmtn_Q1y98bH6RwkHVTM650sZQLPafrFRg/edit?usp=sharing
| + | |
| − | </a>
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | (Just a quick conclusion) Finished the assay and noticed that only Mike’s pDawn + lacZ turned yellow after adding WS; our ligation was not
| + | |
| − | successful
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | But, when we tried to make the standard curve, we got a graph that didn’t make any sense.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | We think it could be because:
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | The equation includes OD660 for something that’s not a cell?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | We waited too long to add the stop solution (too yellow)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | We diluted many times to get the OD420 in the range of 0.2 - 1.0; we may have not mixed the solutions enough (the ODs were all pretty
| + | |
| − | much 0.6)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <h1 dir="ltr">
| + | |
| − | Week 5 (6/29 - 7/3)
| + | |
| − | </h1>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 6/29/15
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims For Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish presentation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | DIscuss project ideas; goals and data with advisors during Monday Meeting
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Figure out trajectory for the summer.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish presentation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | DIscuss project ideas; goals and data with advisors during Monday Meeting
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Figure out trajectory for the summer.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol> | + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Co transform pDawn + lacZ + luxbox (fail-fast)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Redo standard curve for B- Gal assay
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Design experiment for blue-light intensity for assay
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Did lux box + pACBB ligate properly?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Decide which parts to order from IDT (but actually)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Talk about different color circuits
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 6/30/15
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims For Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | grow up pACBB-lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | dilute B-Gal
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up BL21 lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up lux for spiked arabinose experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Fill out safety form
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Set up incubator for blue light vs. dark pDawn experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Get primers
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Sequence pACBB-lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculate pDawn-lacZ for blue light inducibility experiment (dark)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Set up sequencing tubes for pACBB-lux part
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | do colony PCR at pACBB-lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | redo B-gal assay standardized curve
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up BL21 lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up pACBB-lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up lux for spiked arabinose experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Fill out safety form
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Set up incubator for the blue light vs. dark pDawn experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Get primers
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Set up sequencing tubes for pACBB-lux part
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | back dilute pDawn - lacZ 1:100 (dark)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | at OD of around .4, separate→ 3 blue; 3 dark
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | miniprep pACBB lux (2 tubes)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Sequence miniprepped pACBB-lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | PCR test of miniprepped pACBB-lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Redo B-gal assay standardized curve
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Redesign the illuminated incubator
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Friday:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | After 20 h of growth, do OD600 and miller assay for pDawn lacZ light vs. dark
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/1/15
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave flasks
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Set up blue light incubator
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At OD of .4, induce pDawn-lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Back dilute pDawn-lacZ 1:100 (dark) in 25 mL LB Kan
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Dilute B-Gal for curve
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Do B-Gal curve w/ BL21
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep pACBB-lux (4 tubes)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Back-dilute 1:100 lux BL21 for arabinose experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Induce lux w/ arabinose every hour; measure OD + luminescence
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave flasks
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Colony PCR
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Sequencing.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave flasks
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Set up blue light incubator
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At OD of .4, induce pDawn-lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Back dilute pDawn-lacZ 1:100 (dark) in 25 mL LB Kan
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Dilute B-Gal for curve
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Do B-Gal curve w/ BL21
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep pACBB-lux (4 tubes)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Back-dilute 1:100 lux BL21 for arabinose experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Induce lux w/ arabinose every hour; measure OD + luminescence
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave flasks
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Colony PCR
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run PCR of pACBB lux on gel
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pour gel
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Send in sequencing of pACBB-lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At ll AM, miller assay of pDawn light vs. dark
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | If pACBB gel good, do pACBB vs. pSB1C3 experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/2/15
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Sequencing of pACBB-lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pour gel
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run pACBB on gel
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At 11 AM, Miller Assay of pDawn light vs. dark
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Co-culture pSB1C3 + lux and pDawn + lacZ. You have to cotransform the opposite antibiotic resistance into each plasmid before transformation.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Dialysis Tube (ask Mike for this) _ controls for them growing at different rates
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Just pipet them into the same tube
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up a giant culture of pSB1C3 + lux; induce at the right times for visible luminescence
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | reach out to the iGEM teams who did similar experiments to see what they did for a visible glow.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Sequencing of pACBB-lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pour gel
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run pACBB on gel
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At 11 AM, Miller Assay of pDawn light vs. dark
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run PCR of pACBB-lux on gel
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pour gel
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Send in sequencing of pACBB-lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At ll AM, miller assay of pDawn light vs. dark
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | If pACBB gel good, do pACBB vs. pSB1C3 experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <h1 dir="ltr">
| + | |
| − | Week 6 (7/6 - 7/10)
| + | |
| − | </h1>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/6/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave flasks
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miller assay of samples from light vs. dark experiment (samples were stored in the fridge for 4 days, so if it doesn’t work, restart experiment)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Send in sequencing
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 pDawn-lacZ in 25 mL LB-Kan
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Redo PCR of lux-pACBB; troubleshoot?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculate pACBB-lux from glycerol stock
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At an OD of 0.3, induce with arabinose measure luminescence every hour (controls: uninduced pACBB-lux and pACBB-eGFP)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At an OD of 0.3, induce with blue light
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze arabinose spiking data.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave flasks
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miller assay of samples from light vs. dark experiment attempted_ experiment restarted
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Send in sequencing
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 pDawn-lacZ in 25 mL LB-Kan
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Redo PCR of lux-pACBB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculate pACBB-lux from glycerol stock
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At an OD of 0.3, induce with arabinose measure luminescence every hour (controls: uninduced pACBB-lux and pACBB-eGFP)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At an OD of 0.3, induce with blue light
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze arabinose spiking data.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | > Debrief from monday meeting
| + | |
| − | </p>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | sequence Mike’s pDawn+lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | research lacZ degradation rate (decide if we need to characterize)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | if we can’t find much, expose pDawn lacZ to blue light for 2h then measure lacZ expression every 2 hours until 20 h
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | clone pDawn essentials onto pACBB backbone (make primers to PCR off pDawn portion needed)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | adjust beta-galactosidase curve for concentration
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | discuss Karol database stuff (we disagree with Monday’s discussion)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | be more careful with what we put in the Monday meeting slide
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | don’t put in tentative data
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | keep it short
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | focus on the questions you have
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | focus on graphs, experimental designs, conclusions
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | data into excel spreadsheet
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | talk to Victor about lacZ, decide whether it’s worth it to maybe switch to GFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | ask Victor about how quickly it degrades
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | as soon as sequencing results come back, cotransform
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | either with pACBB or other compatible plasmids with the opposite antibiotic resistance
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | do an experiment with
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | do the poster 2 weeks before the July 31st deadline
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | present it at the meeting, so we can nip any potential problems in the butt
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | talk about restructuring our day and integrating most poster/website stuff
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | include laying out experimental designs and sending them out
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | </ul>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Questions:
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/7/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miller assay
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Arabinose spiking experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Research on lacZ degradation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pour gel
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up from glycerol stock *B2* BL21 lux for OD experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run gel of pACBB PCR
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Talk to Seville about equipment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Sequence pDawn+lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plan lacZ experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Look into XGal
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow Mike’s pDawn *get from Mike
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Figure out luminometer
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up pDawn-LacZ (A1)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miller assay
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pour gel
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run gel of pACBB PCR
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plan lacZ-degrading experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 for OD experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Induce with arabinose at dif OD and take luminescence measurements
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Illuminometer meeting with Mike
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 of pDawn-lacZ in 25 mL
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At OD of 0.4, induce with blue light
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Measure lacZ every 3 hours
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/8/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Sequence rest of pACBB-lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Sequence pDawn-lacZ (Mike’s) with primers that bind to outside
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pour gel
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run gel of pDawn+lacZ PCR
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Do pDawn+mRFP light vs. dark experiment (1:100 at 11 AM and induce at 2 PM)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 BL21 lux in 25 mL LB+chlorom
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Design primers to swap out promoter in lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB vs. pSBIC3 experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Backdiluted BL21 and DH5-alpha with pDawn RFP 1:100 for blue vs. dark experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Induced pDawn RFP strains at OD ~.3-.4 with blue light
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Poured gel, ran gel of pDawn lacZ PCR check → success! lacZ amplified, next step if to sequence pDawn lacZ to make sure the band is what we think
| + | |
| − | it is
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculated BL21 lux from glycerol stock for arabinose induction at different OD experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Designed primers to change lux operon promoter to T7 (not pBAD) by cloning into pRSFDuet and pET-26b(+) backbones
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Backdiluted pACBB lux 1:100, induced with arabinose at OD ~.4, took OD measurements every hour for 7h (to characterize pACBB lux expression vs.
| + | |
| − | pSCB1C3 lux expression)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Backdilute BL21 lux 1:100 in LB Chlor (9 AM)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Measure RFP expression in pDawn blue light vs. dark experiment at 20 h (10 AM)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Backdilute pDawn lacZ 1:100 in 25 mL LB Kan to redo blue vs. dark experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | induce pDawn lacZ with blue light at OD ~.4
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep pDawn lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Send of pDawn lacZ for sequencing with primers (2) that bind to outside pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze pACBB vs. pSB1C3 luminescence results
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Think about swapping out pDawn backbone for pACBB and redoing lacZ cloning (decide what to do moving forward)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Questions:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Should we move forward with changing the lux operon promoter to T7 ASAP, and should we order the primers for the cloning tomorrow?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/9/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock pDawn RFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Induce pDawn lacZ at OD .4 for light vs dark experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprepped pDawn RFP and pACBB lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Induce BL21 with arabinose at different OD and measured luminescence
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Sequence miniprepped pDawn lacZ with pDawn binding primers
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Sequence miniprepped pACBB lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze pACBB vs. PSB1C3 lux results
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Think about swapping out pDawn backbone
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze pDawn RFP light vs. dark results
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Cotransformed pDawn RFP and pACBB lux into BL21
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock pDawn RFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprepped pDawn RFP and pACBB lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyzed pACBB vs. PSB1C3 lux results
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyzed pDawn RFP blue light vs. dark results
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Looked into h-ns mutant E. coli and effects on lux operon expression
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Looked into other plasmids that would luminesce better than the lux operon biobrick
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Backdilute BL21 lux 1:100 in 40 mL LB Chlor for OD arabinose induction experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Sequence miniprepped pDawn lacZ with pDawn binding primers
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Sequence miniprepped pACBB lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Think about swapping out pDawn backbone
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Induce BL21 with arabinose at different OD and measured luminescence (started today but issues with Tecan readings)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pick colonies from transformation plates, inoculate, glycerol stock (glycerol stock later in day)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Look into ordering h-ns mutant E. coli and ordering plasmids off Addgene that may luminesce brighter than lux operon BioBrick
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Questions:
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/10/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Back dilute BL21 lux 1:100 for temperature experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Send off pDawn lacZ, pDawn RFP, and pACBB lux for sequencing
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | induce BL21 with arabinose and grow at different temperature
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pick colonies from cotransformation of pACBB lux and pDawn RPF
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Set up experiment to see if pACBB lux induced RPF expression
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | organize data
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | prepare BL21 competent cells
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | glycerol stock pDawn lacZ/RFP + pACBB lux BL21 strains from cotransformation (4 colonies per cotransformation)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Back dilute BL21 lux 1:100 for temperature experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Send off pDawn lacZ, pDawn RFP, and pACBB lux for sequencing
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Induce BL21 with arabinose and grow at different temperature, measure every hour
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pick colonies from cotransformation of pACBB lux and pDawn RPF
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Set up experiment to see if pACBB lux induced RPF expression
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finalize what to spend the 20kb on from IDT
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order primers for changing lux operon promoter
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculate pDawn RFP + pACBB lux colonies for fail fast attempt number two, backdilute 1:100 after grown to saturation, induce with arabinose at OD
| + | |
| − | ~.4
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | After 20h, measure RFP expression for all fail fast tubes (Tuesday)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze lux and pDawn sequencing results
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Email asking for the split lux plasmid from this (
| + | |
| − | <a href="http://link.springer.com/article/10.1007/s00216-010-4266-7/fulltext.html">
| + | |
| − | http://link.springer.com/article/10.1007/s00216-010-4266-7/fulltext.html
| + | |
| − | </a>
| + | |
| − | ); specifically asking for Sy102 and SY104
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Cotransform opposite antibiotic resistence for coculturing experiment (pUC57 Kan, find something for pDawn RFP)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Prepare BL21 competent cells (grow up cells in the morning to OD .3-.4 is the first step)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Questions:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | How do you make BL21 competent cells?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | What white microwell plates should we order?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Can we order primers to move ahead with replacing the pBAD promoter in lux with a T7 promoter?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | How should we plan the fail fast experiment? Right now, we have it set up so each cotransformed colony will grow in 1 tube with light (not induced
| + | |
| − | with arabinose to check for pDawn presence/activity alone) and two tubes in the dark (one induced with arabinose as our “test” tubes and one
| + | |
| − | uninduced to show that pDawn is not induced without lux operon expression). All of these are done in 1:1 LB kan LB Chlor media. We also have a
| + | |
| − | negative control of BL21 with the lux operon alone growing in LB kan+chlor to show that it will not grow. This tube will remain in the light and
| + | |
| − | idk if we should induce it with arabinose or not. Anyway, are there any other controls we should seriously consider including?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <h1 dir="ltr">
| + | |
| − | Week 7 (7/13 - 7/17)
| + | |
| − | </h1>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/13/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plan RFP-lux experiment #2
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Innoculate from glycerol stock
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plan Tecan calibration experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Ask Mike + Spencer to grant us access to BE undergrad lab
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze sequencing for pDawn lacZ and lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Innoculate from glycerol stock
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plan Tecan calibration experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Prepare presentation.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/14/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave flasks
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make BL21 competent cells (6; grow up 1)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make LB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make LB + chlor
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Co transform w/ opposite antibiotic
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Ask Mike for pet26B
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order primers to swap out GFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Tecan test
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | inoculate A5 and B2/B3
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Print a toucan
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 pDawn + lacZ; At an OD of 0.4 induce
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 cotransformed stuff; At an OD of 0.4 induce
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make arabinose
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order luminol + email iGEM team
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze sequencing results.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave flasks
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze sequencing results.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make arabinose
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 cotransformed stuff; At an OD of 0.4 induce
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculate
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Tecan test-
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | A1 - Mike’s ligated pDawn + lacZ (as a negative control)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | B2 - pSB1C3 + lux box (to test luminescence)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miller Assay (light vs. dark)-
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | A1 - Mike’s ligated pDawn + lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | B2 - pSB1C3 + lux box (negative control)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | BL21 + lux - USE B2
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Mike’s ligated pDawn +lacZ - USE A1
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux - USE A7
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pick up LB from the autoclave
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make BL21 competent cells
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make BL21 glycerol stock
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Tecan test to make sure that luminescence measurements are correct
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Co transform w/ opposite antibiotic
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Ask Mike for pet26B
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order primers to swap out GFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 pDawn + lacZ.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Induce with blue light at an OD of 0.4
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculate cotransformed pACBB + lux box & pDawn + lacZ for experiment tomorrow.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze data for cotransformed mRFP experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | START POSTER
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Thursday:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run two experiments simultaneously
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Nikita/Hannah
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miller assay for pDawn + lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | After 20 hours, measure lacZ expression
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze data
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Jane/Karol
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Repeat cotransformed mRFP experiment with cotransformed lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Based on results from the first experiment, validate the second experiment.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/15/15
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make new LB (since we messed it up)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make BL21 competent cells
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make BL21 glycerol stock
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up pet26B
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 pDawn + lacZ.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Induce with blue light at an OD of 0.4
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculate cotransformed pACBB + lux box & pDawn + lacZ for experiment tomorrow.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze data for cotransformed mRFP experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make new LB (since we messed it up) + LB chlor
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up pet26B
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 pDawn + lacZ.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Induce with blue light at an OD of 0.4
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculate cotransformed pACBB + lux box & pDawn + lacZ for experiment tomorrow.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze data for cotransformed mRFP experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pla big flask culture experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 20 hr of pDawn-lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 20 hr of RFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run big flask culture experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze RFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn-LacZ Failfast Experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At 9AM do a 1:100 dilution
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Set up PET primers
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Design high GC-lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Email Cambridge iGEM
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Black well plates
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order luminol
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order primers to swap out GFP and RFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/16/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 20 hr of pDawn-lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 20 hr of RFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run big flask culture experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze RFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn-LacZ Failfast Experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At 9AM do a 1:100 dilution
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Set up PET primers
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Design high GC-lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Email Cambridge iGEM
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Black well plates
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order luminol
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order primers to swap out GFP and RFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 20 hr of pDawn-lacZ light vs. dark experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 20 hr of RFP reading
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run big flask culture experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze RFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn-LacZ Failfast Experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At 9AM do a 1:100 dilution
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miller assay at 3 time points (2,5 and 7 hours)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | OD and luminescence readings every hour
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Set up PET primers
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Email Cambridge iGEM
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order black well plates
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze data from pDawn-lacZ failfast experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze data from pDawn-RFP failfast experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Determine if the failfast worked
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Start the Monday Meeting presentation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Talk about final goals
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/17/15
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze data from pDawn-lacZ failfast experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze data from pDawn-RFP failfast experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Determine if the failfast worked
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Start the Monday Meeting presentation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Talk about final goals
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze data from pDawn-lacZ failfast experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze data from pDawn-RFP failfast experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Determine if the failfast worked
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Start the Monday Meeting presentation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Talk about final goals
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Monday:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Determine direction
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish Monday Meeting presentation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <h1 dir="ltr">
| + | |
| − | Week 8 (7/20 - 7/24)
| + | |
| − | </h1>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/20/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today/Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish analyzing data
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Prepare for Monday Meeting presentation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Talk more about our goals and direction now that the failfast didn’t “work”
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | How do we get it to glow?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform pACBB+lux and pSB1C3 + lux in Top10/ NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make the photon counter <- HIGHEST PRIORITY
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order the inverter
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order T7 promoters
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Email Goulian about H-NS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Email about split lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Call Glowell
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order high GC lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/21/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform pACBB+lux and pSB1C3 + lux in Top10/ NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make the photon counter
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order the inverter
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order T7 promoters
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Email Goulian about H-NS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Email about split lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Call Glowell
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order high GC lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make the photon counter
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order T7 promoters
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Email about split lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Call Glowell
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order high GC lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform pACBB+lux and pSB1C3 + lux in Top10/ NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish the photon counter
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order the inverter
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Email Goulian about H-NS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/22/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish the photon counter
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Email Goulian about H-NS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Poster deadline is approaching
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Work on the presentation for Seville’s talk
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make the photon counter
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Heard back about split lux: going to send it in two weeks if we want it
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Emailed Goulian and the lab Cambridge got their hns mutant strains from
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform into Top10/ NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 OF NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow to an OD of 0.4
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Spin them down
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pour out the LB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Resuspend in TSS buffer ← look in the Chow lab
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | NEB Turbo (- control)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | B9, B1, B4, G2 and cotransform G2 and F4
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Next week
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Leave plates out on the counter? On Monday, pick from them and ultimately make glycerol stock
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Redo lux experiments
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/22/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol start="6">
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish the photon counter
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Email Goulian about H-NS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Poster deadline is approaching
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Work on the presentation for Seville’s talk
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol start="4">
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make the photon counter
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Heard back about split lux: going to send it in two weeks if we want it
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Emailed Goulian and the lab Cambridge got their hns mutant strains from
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol start="2">
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform into Top10/ NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 OF NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow to an OD of 0.4
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Spin them down
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pour out the LB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Resuspend in TSS buffer ← look in the Chow lab
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | NEB Turbo (- control)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | B9, B1, B4, G2 and cotransform G2 and F4
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Next week
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Leave plates out on the counter? On Monday, pick from them and ultimately make glycerol stock
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Redo lux experiments
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/23/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | -Skip-
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/24/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make NEB10 competent
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 OF NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow to an OD of 0.4
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Spin them down
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pour out the LB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Resuspend in TSS buffer ← look in the Chow lab
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish working on the photon counter experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pick up primers from cell center
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish sending emails to source parts
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make NEB10 competent
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 OF NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow to an OD of 0.4
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Spin them down
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pour out the LB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Resuspend in TSS buffer ← look in the Chow lab
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish working on the photon counter experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol start="3">
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform into Top10/ NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | NEB Turbo (- control)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | B9, B1, B4, G2 and cotransform G2 and F4
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pSB1C3 + lux (B9)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | miniprepped pSB1C3 + lux (B1)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux (B4)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn + RFP (G2)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <br/>
| + | |
| − | <ol start="2">
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Next week
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Leave plates out on the counter? On Monday, pick from them and ultimately make glycerol stock
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Redo lux experiments
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <h1 dir="ltr">
| + | |
| − | Week 9 (7/27 - 7/31)
| + | |
| − | </h1>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/27/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform parts into NEB10 Competent Cells
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | B9, B1, B4, G2 and cotransform G2 and F4
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | NEB Turbo (- control)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pSB1C3 + lux (B9)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | miniprepped pSB1C3 + lux (B1)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux (B4)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn + RFP (G2)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish Monday Meeting presentation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finalize information for the website/poster
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plan experiment to measure luminescence with NEB10 tomorrow.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | “Big-ass culture” -->3 g/ 20 mL arab
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish photon counter (determine fate?)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Call Tecan
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Ask Jane
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | T7_pickup cell center
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Codon optimized
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inverter strain
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Send Fedex for Split Lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol start="8">
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform parts into NEB10 Competent Cells
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | B9, B1, B4, G2 and cotransform G2 and F4
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | NEB Turbo (- control)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pSB1C3 + lux (B9)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | miniprepped pSB1C3 + lux (B1)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux (B4)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn + RFP (G2)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish Monday Meeting presentation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finalize information for the website/poster
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plan experiment to measure luminescence with NEB10 tomorrow.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | “Big-ass culture” -->3 g/ 20 mL arab
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Determine fate of photon counter
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Call Tecan
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Ask Jane
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | T7_pickup cell center
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Codon optimized
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inverter strain
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Emailed Yolanda about Fedex for Split Lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plan out entire summer
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plan future experiments
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/28/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Flesh out goals for toolbox “story”
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform BBa_K525998 T7 + RBS Biobrick into competent cells (lux gene will be transformed into it to make pSB1C3 + lux)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Innoculate cultures
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn + mRFP in NEB10 (F4 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in NEB10 (G2 dna) X 4 (three from same colony to run big ass culture) ::::: (the other is to eventually use for pCR of part)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pSB1C3 + lux in NEB10 (B2 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprepped pSB1C3 + lux in NEB10 (B9 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Cotransformed (pDawn + mFRP + pACBB + lux) in NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in BL21 (DONE)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Obtain HNS strain form Goulian
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Flesh out goals for toolbox “story”
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform BBa_K525998 T7 + RBS Biobrick into competent cells (lux gene will be transformed into it to make pSB1C3 + lux)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Innoculate cultures
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn + mRFP in NEB10 (F4 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in NEB10 (G2 dna) X 4 (three from same colony to run big ass culture) ::::: (the other is to eventually use for pCR of part)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pSB1C3 + lux in NEB10 (B2 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprepped pSB1C3 + lux in NEB10 (B9 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Cotransformed (pDawn + mFRP + pACBB + lux) in NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in BL21 (DONE)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Obtain HNS strain form Goulian
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Figure out how to make T7 + pACBB + lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in NEB10 big ass culture to test for visible luminescence
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn + mRFP in NEB10 (F4 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in NEB10 (G2 dna) X 4 (three from same colony to run big ass culture) ::::: (the other is to eventually use for pCR of part)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pSB1C3 + lux in NEB10 (B2 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprepped pSB1C3 + lux in NEB10 (B9 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Cotransformed (pDawn + mFRP + pACBB + lux) in NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pick
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | HNS that Goulian gave us
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | BBa_K525998 T7 + RBS Biobrick
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Research
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | If we can use the T7 promoter in the HNS strain
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | If the HNS strain mutant (for AraD) so that the arabinose is not eaten up
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Creation of T7 + pSB1C3 + lux system
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep pACBB + lux in BL21 to later be used for pCR
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep BBa_K525998 T7 + RBS Biobrick
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plan for digestion, ligation and transformation on Thursday
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Give presentation to PFP scholars
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up pACBB + lux and cotransformed pACBB + lux & pDawn + mRFP for fail fast experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | TOOLBOX STORY → bolded combinations are prepared, but not tested. Italicized combinations are completed; nothing has been tested with nonanol; yellow are
| + | |
| − | knocked out because T7 promoter does not function in NEB10
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Possible combinations: TOOLBOX 1 LUMINESCENCE
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + T7 + high pSB1C3 + BL21 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + T7 + high pSB1C3 + NEB10 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + T7 + high pSB1C3 + HNS (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + T7 + low pACBB + BL21 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + T7 + low pACBB + NEB10 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + T7 + low pACBB + HNS (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + pBAD + high pSB1C3 + BL21 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + pBAD + high pSB1C3 + NEB10 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + pBAD + high pSB1C3 + HNS (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + pBAD + low pACBB + BL21 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + pBAD + low pACBB + NEB10 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + pBAD + low pACBB + HNS (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Possible combinations: TOOLBOX 2 SENDER RECEIVER SYSTEM (Simultaneously done)
| + | |
| − | </p>
| + | |
| − | <ol start="4">
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + T7 + high pSB1C3 + BL21 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + T7 + high pSB1C3 + NEB10 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + T7 + high pSB1C3 + HNS (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <ol start="4">
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + T7 + low pACBB + BL21 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + T7 + low pACBB + NEB10 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + T7 + low pACBB + HNS (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <ol start="4">
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + pBAD + high pSB1C3 + BL21 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + pBAD + high pSB1C3 + NEB10 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + pBAD + high pSB1C3 + HNS (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <ol start="4">
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + pBAD + low pACBB + BL21 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + pBAD + low pACBB + NEB10 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + pBAD + low pACBB + HNS (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | IF TIME, TOOLBOX 2
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux AB + CDE
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux AB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux CDE
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | HUMAN PRACTICES
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Two inverter strains
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/29/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Figure out how to make T7 + pACBB + lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in NEB10 big ass culture to test for visible luminescence
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn + mRFP in NEB10 (F4 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in NEB10 (G2 dna) X 4 (three from same colony to run big ass culture) ::::: (the other is to eventually use for pCR of part)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pSB1C3 + lux in NEB10 (B2 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprepped pSB1C3 + lux in NEB10 (B9 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Cotransformed (pDawn + mFRP + pACBB + lux) in NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pick
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | HNS that Goulian gave us
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | BBa_K525998 T7 + RBS Biobrick
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Research
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | If we can use the T7 promoter in the HNS strain
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | If the HNS strain mutant (for AraD) so that the arabinose is not eaten up
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Creation of T7 + pSB1C3 + lux system
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep pACBB + lux in BL21 to later be used for pCR
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep BBa_K525998 T7 + RBS Biobrick
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plan for digestion, ligation and transformation on Thursday
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Give presentation to PFP scholars
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up pACBB + lux and cotransformed pACBB + lux & pDawn + mRFP for fail fast experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Figure out how to make T7 + pACBB + lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plan for pACBB + lux in NEB10 big ass culture to test for visible luminescence (there weren’t enough flasks to do it yesterday)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave flasks
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in NEB10 (G2 dna) X 4 (three from same colony to run big ass culture) ::::: (the other is to eventually use for pCR of part)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pSB1C3 + lux in NEB10 (B2 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprepped pSB1C3 + lux in NEB10 (B9 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Cotransformed (pDawn + mFRP + pACBB + lux) in NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pick
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | HNS that Goulian gave us
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | BBa_K525998 T7 + RBS Biobrick
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn + mRFP in NEB10 (F4 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Research
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | If we can use the T7 promoter in the HNS strain
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | If the HNS strain mutant (for AraD) so that the arabinose is not eaten up
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Creation of T7 + pSB1C3 + lux system
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep pACBB + lux in BL21 to later be used for pCR
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plan for digestion, ligation and transformation on Thursday
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow::
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish the poster
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in NEB10 in big ass culture
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep BBa_K525998 T7 + RBS Biobrick
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 HNS at 11AM
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make HNS cells competent
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol Stock pDawn + mRFP in NEB10 (F4 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Cleanup pCR product (lux)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digestion of pCRed lux + miniprepped Biobrick
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform HNS with pBAD + pACBB + lux & pBAD + pSB1C3 + lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Overnight ligation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/30/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish the poster
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in NEB10 in big ass culture
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep BBa_K525998 T7 + RBS Biobrick
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Cleanup pCR product (lux)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digestion of pCRed lux + miniprepped Biobrick
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Overnight ligation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 HNS at 11AM
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make HNS cells competent
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up HNS for big culture tomorrow
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock HNS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform HNS with pBAD + pACBB + lux & pBAD + pSB1C3 + lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol Stock pDawn + mRFP in NEB10 (F4 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave BE lab flasks and flasks for big ass HNS culture tomorrow
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Bleach everything on the table in the back room
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Talk more about switching out the promoter; ask the advisors if we should bother choosing
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in NEB10 in big ass culture
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep BBa_K525998 T7 + RBS Biobrick
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Cleanup pCR product (lux)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digestion of pCRed lux + miniprepped Biobrick + cleanup
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 HNS at 11AM
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make HNS cells competent
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up HNS for big culture tomorrow
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock HNS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform HNS with pBAD + pACBB + lux & pBAD + pSB1C3 + lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol Stock pDawn + mRFP in NEB10 (F4 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave BE lab flasks and flasks for big ass HNS culture tomorrow
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Bleach everything on the table in the back room
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Talk more about switching out the promoter; ask the advisors if we should bother choosing
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digestion + Cleanup
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Overnight ligation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Big ass culture for HNS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock HNS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform HNS with pBAD + pACBB + lux & pBAD + pSB1C3 + lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave BE lab flasks
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Talk more about what final goals will be
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish poster
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Figure out luminometer
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | If time, transform T7 promoter construct into BL21; also cotransform with pDawn + mRFP (GROW UP IN THE MORNING)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 cotransformed cultures
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Measure OD, luminescence and RFP every hour
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Questions:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Should we use the T7 promoter if we cannot put it into HNS or NEB10? Does it detract from the comprehensiveness of the toolbox?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/31/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digestion + Cleanup
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Overnight ligation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Big ass culture for HNS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock HNS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform HNS with pBAD + pACBB + lux & pBAD + pSB1C3 + lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave BE lab flasks
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Talk more about what final goals will be
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish poster
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Figure out luminometer
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | If time, transform T7 promoter construct into BL21; also cotransform with pDawn + mRFP (GROW UP IN THE MORNING)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 cotransformed cultures
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Measure OD, luminescence and RFP every hour
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digestion + Cleanup
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock HNS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform HNS with pBAD + pACBB + lux, pBAD + pSB1C3 + lux, pBAD + pACBB, pDawn + mRFP and cotransformed pACBB + lux and pDawn + mRFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave BE lab flasks
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Complete poster (or just generate an idea to be completed on the weekend)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Monday:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Do the cotransformed mRFP experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 cotransformed cultures
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Measure OD, luminescence and RFP every hour
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Talk more about final goals
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Iron out the experiments that we have planned for the rest of the summer
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | tracking
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | proving orthogonality
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Prepare final poster presentation to send out to the advisors on Sunday night
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Prepare for Tuesday Advisor Meeting
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <h1 dir="ltr">
| + | |
| − | Week 10 (8/3 - 8/7)
| + | |
| − | </h1>
| + | |
| − | <p dir="ltr">
| + | |
| − | 8/3/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Do the cotransformed mRFP experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 cotransformed cultures
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Measure OD, luminescence and RFP every hour
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Talk more about final goals
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Iron out the experiments that we have planned for the rest of the summer
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | tracking
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | proving orthogonality
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Prepare final poster presentation to send out to the advisors on Sunday night
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Prepare for Tuesday Advisor Meeting
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Talk more about final goals
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Iron out the experiments that we have planned for the rest of the summer
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Prepare for Tuesday Advisor Meeting
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Right now
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Orthogonality→ proving for sure that light and chemicals are orthogonal to each other
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Tracking → chemical vs. light
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | This experiment will demonstrate hopefully that light tracks better than chemicals which would mean it has better spatiotemporal
| + | |
| − | control
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | These two principles can be used to build a major experiment (Pim’s idea)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | A bidirectional communication system in which one cell communicates with the second cell with chemicals and the same second cell
| + | |
| − | communicates with the first using light. This should create a “stable” system which oscillates around a line because the K2 (light
| + | |
| − | communication) is higher than K1 (chemical communication). It is a form of negative feedback.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make adjustments to the presentation for advisor meeting with Dr. Chow
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Iron out more of the cloning details for Pim’s bidirectional idea
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make changes based on advisor feedback
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 8/4/15
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make adjustments to the presentation for advisor meeting with Dr. Chow
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Iron out more of the cloning details for Pim’s bidirectional idea ← Too complicated; not necessarily based on an engineering principle; our
| + | |
| − | abstract talks about this bidirectional communication, but our experiments are not necessarily aligning with it.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Rework the presentation to fit a more probable system that we can use.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | A semi-completed poster
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | What we thought was bistable communication, but instead was just a system that would permanently exist in the second state after addition of
| + | |
| − | light
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Prepare a distance experiment in which you have six tubes and place tubes inside them so that you can run “co-tubing experiment”
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up NEB10 glycerol stock one in the dark (pDawn + mRFP) and one in the light (pACBB + lux)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 dilute into flasks after reaching saturation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Measure at 20 hours after separating into individual tubes
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Adjust experiments based on if you can get this to work
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Prepare a list of principles that we want to base our entire project on; come up with a list of experiments that complement this list
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Ideas discussed during the meeting:
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pim’s tracking idea in which we mutate the lov domain to create different tracking curves
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Possibly a distance idea→ further iron out the application of this type of a project
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 8/5/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Prepare a distance experiment in which you have six tubes and place tubes inside them so that you can run “co-tubing experiment”
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up NEB10 glycerol stock one in the dark (pDawn + mRFP) and one in the light (pACBB + lux)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 dilute into flasks after reaching saturation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Measure at 20 hours after separating into individual tubes
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Adjust experiments based on if you can get this to work
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Prepare a list of principles that we want to base our entire project on; come up with a list of experiments that complement this list
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Prepare a list of principles that we want to base our entire project on; come up with a list of experiments that complement this list
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Current ideas on the table
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Light compatible with strains or environment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Tuneable photoreceptors with the lov mutation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Luminescence signal → less persistent signal
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Chemical and light → different rates of production; orthogonality
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Light driven spatiotemporal control of chemical communication
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Ironed out specific experiments for the above
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Prepared the “co-tubing” experiment to run tomorrow
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Go through the principles/project ideas with advisors and narrow down to the ones we want
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish the co-tubing experiments and find out which ones are feasible ^
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish cloning the T7 promoter
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Characterize individual sender + receiver parts
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Find out what the sender cell curve looks like in NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Find out if the receiver responds to the sender cell; simulate the light curve to test this???
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plan experiments for the chosen project idea more thoroughly
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Redo poster for next Monday Meeting with Dr. Chow
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 8/6/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish ranking the projects/principles
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Take flasks out of autoclave at 12 PM
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 dilute them
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Induce at OD of 0.3
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Take 20 hr measurement tomorrow
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish pACBB + lux in NEB10 luminescence curve
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inverter
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Identify 3 with different strengths
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Email advisors about projects; remind them that the abstract is due tomorrow
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Email advisors about projects; remind them that the abstract is due tomorrow
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish cloning T7
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Retransform into HNS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | F4
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | B9
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | G2
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | F4 + G2
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <h1 dir="ltr">
| + | |
| − | -Skip Week 11-
| + | |
| − | </h1>
| + | |
| − | <h1 dir="ltr">
| + | |
| − | Week 12 (8/17 - 8/21)
| + | |
| − | </h1>
| + | |
| − | <p dir="ltr">
| + | |
| − | 8/17/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | inoculate pdawn rfp (dh5alpha and BL21) for receiver characterization
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | inoculate psb1c3 lux in NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | backdilute psb1c3 lux at 6 PM
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | induced lux psb1c3 with arabinose at 10 PM (when OD at ~.4)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | order LB plates
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | finish soldering circuit for pDawn receiver characterization
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | setup tubes for pDawn receiver characterization experiment (to start tomorrow) (program arduino to turn off blue light at certain times)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | plate agar stab with split lux operon
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | plate agar stabs with each antibiotic only to separate plasmids
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | transform psb1c3 lux operon into h-ns
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | regraph pelleting volume luminescence data with photons on x-axis
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Publication for reference on split lux: http://link.springer.com/article/10.1007/s00216-010-4266-7/fulltext.html
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | inoculate pdawn rfp (dh5alpha and BL21) for receiver characterization
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | inoculate psb1c3 lux in NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | backdilute psb1c3 lux at 6 PM
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | induced lux psb1c3 with arabinose at 10 PM (when OD at ~.4)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | order LB plates
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | finish soldering circuit for pDawn receiver characterization
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | plate agar stab with split lux operon
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | plate agar stabs with each antibiotic only to separate plasmids
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | transform psb1c3 lux operon into h-ns
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tuesday:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | backdilute pDawn RFP 1:100 for receiver characterization
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | induce pDawn RFP with light at 10 AM (when OD at ~.4)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | turn off blue light at appropriate times for pdawn receiver characterization (or check that arduino is working)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 8 AM → take a 10 h timepoint for luminescence of psb1c3 lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | take hourly luminescence and OD readings for psb1c3 lux up to 17 h (3PM)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pick split lux operon colonies, inoculate (at least 3 colonies)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pick and replate colonies for attending to separate out plasmids
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pick h-ns lux transformed colonies, inoculate
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 8/18/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | glycerol stock h-ns transformed with psb1c3 lux (at least 3 colonies)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | glycerol stock split lux operon colonies (at least 3 colonies)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | inoculate h-ns lux from glycerol stock for time course experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pick and replate colonies for attending to separate out plasmids for second time
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Replan NEB10 pSB1C3 + lux/ HNS pSB1C3 + lux experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Website
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make LB Media
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | chlor
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | amp
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | chlor + amp
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | kan
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | To Make Glycerol stocks:
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up SY102 in Chlorom to isolate CDE
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up SY104 in Chlorom to isolate CDE
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up SY102 in Amp to isolate AB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up SY102 in Amp + Chlorom to isolate split lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up SY104 in Amp + Chlorom to isolate split lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | To see if plasmid has been lost
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up SY102 plated on Chlor in Amp to see if it survives
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up SY104 plated on Chlor in Amp to see if it survives
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up SY102 plated on Amp in Chlor to see if it survives
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | </ul>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | For the timecourse experiment:
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Take the following upstairs:
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pSB1C3 + lux in HNS (x2)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pSB1C3 + lux in NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in HNS (x2)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn + mRFP in BL21 → should be in foil
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn + mRFP in DH5alpha→ should be in foil
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Take aluminum foil upstairs just in case you cannot find it.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At 9PM,
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Measure OD of cultures below to ensure that they have reached saturation (see if one of the two tubes has not grown properly)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in HNS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in HNS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 in 100 ml media culture flasks (so we add 1000 ul of culture into the appropriate flask)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pSB1C3 + lux in HNS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pSB1C3 + lux in NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in HNS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 in 20 ml media culture flasks(so we add 200 ul of culture into the appropriate flask) COVER GIANT FLASKS IN FOIL
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn + mRFP in BL21 → should be in foil
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn + mRFP in DH5alpha→ should be in foil
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Place all flasks in the incubator; make sure they are closed; make sure that the flasks are not falling out
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At 11:30-12:00PM,
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Measure the OD of ALL cultures; make sure you use conversion to make sure it is appropriate to induce.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Separate the following cultures into three tubes with 5 ml of liquid in each one. Then add 50 ul of arabinose to each tube.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pSB1C3 + lux in HNS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pSB1C3 + lux in NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in HNS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Also add arabinose to the big flasks and place them in teh incubator
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Clean the inside of the culture tube with Karol’s set up with isoproponol. Add 3 mL of the appropriate culture into each tube. Turn on the
| + | |
| − | light by making sure the switch is in - position and that the yellow is connected to red and black is connected to black.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Once the light turns on, cover the lids with aluminum foil and make sure controls are also properly covered.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | </ul>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Sender-cell trendlines:
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Start 10-17 hour luminescence measurement for pSB1C3 + lux in both HNS and NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze the data with the pDawn + mRFP experiment. Determine the threshold?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 (at 10:30) pSB1C3 + lux in HNS (in LB + chlorom) to observe the trend in hours 1-6
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At 12:30 PM
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Separate into 4 different tubes with 3 mL each (1 is an uninduced control)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Add 30 ul of 1 M arabinose to each tube and take OD and luminescence measurements (approximately 1 PM - 7 PM)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Housekeeping:
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock SY102 and SY104 + add to LIMS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pick up plates and nalidixic acid from the cell center.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pour LB + Chlor plates
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plan co-tubing experiment:
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 pSB1C3 + lux in HNS_1000 uL to 100 mL; 1:100 pDawn + mRFP in DH5alpha (in the dark)__500 uL to 50 mL
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Prepare 6 tubes
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Tube 1 (X2)- in dark; IND 10 mL of pSB1C3 + lux; 3 mL of pDawn + mRFP inside
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Tube 2 (X2)- in dark; UNIND 10 mL of pSB1C3 + lux; 3 mL of pDawn + mRFP inside
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Tube 3 (X2)- in light; 3 mL of pDawn + mRFP inside
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At 2 hours, check the OD has reached ~0.3-0.4. Add the appropriate volume of the appropriate culture to the tubes after inducing with 1 M
| + | |
| − | arabinose.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At 20 hr take a fluorescence measurement.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Replating for split lux (yesterday proved that the cells have not lost the plasmid)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Re-plate SY102 in Chlorom to isolate CDE
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Re-plate SY104 in Chlorom to isolate CDE
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Re-plate SY102 in Amp to isolate AB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Tomorrow grow up all plates in opposite antibiotic.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 8/19/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol start="5">
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Sender-cell trendlines:
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Start 10-17 hour luminescence measurement for pSB1C3 + lux in both HNS and NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze the data with the pDawn + mRFP experiment. Determine the threshold?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Housekeeping:
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock SY102 and SY104 + add to LIMS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pick up plates and nalidixic acid from the cell center.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pour LB + Chlor plates
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plan co-tubing experiment:
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 pSB1C3 + lux in HNS_1000 uL to 100 mL; 1:100 pDawn + mRFP in DH5alpha (in the dark)__1000 uL to 100 mL
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Prepare 6 tubes
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Tube 1 (X2)- in dark; IND 10 mL of pSB1C3 + lux; 3 mL of pDawn + mRFP inside
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Tube 2 (X2)- in dark; UNIND 10 mL of pSB1C3 + lux; 3 mL of pDawn + mRFP inside
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Tube 3 (X2)- in light; 3 mL of pDawn + mRFP inside
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At 2 hours, check the OD has reached ~0.3-0.4. Add the appropriate volume of the appropriate culture to the tubes after inducing with 1 M
| + | |
| − | arabinose.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At 20 hr take a fluorescence measurement.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Replating for split lux (yesterday proved that the cells have not lost the plasmid)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Re-plate SY102 in Chlorom to isolate CDE
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Re-plate SY104 in Chlorom to isolate CDE
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Re-plate SY102 in Amp to isolate AB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Tomorrow grow up all plates in opposite antibiotic.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | For Jane,
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | HNS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Split lux (both strains)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make sure she has enough flasks
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 at 6 AM to 7 AM ??? so induce at 8 AM - 9AM
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make sure she has 1 M arabinose and appropriate nalidixic acid solution made up
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Split up the timepoints so that she doesn’t have to do everything by herself
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Website and poster layout completed.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Penn iGEM 2015 Daily Digest
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p>
| + | |
| − |
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | <a href="https://docs.google.com/document/d/1HvOEUNTXplBFVLyiLFqUM917iPTD_HMbZdsruq0erJE/edit#heading=h.sxfktjtcmvnc">Week 2 (6/8 - 6/12)</a>
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | <a href="https://docs.google.com/document/d/1HvOEUNTXplBFVLyiLFqUM917iPTD_HMbZdsruq0erJE/edit#heading=h.bhn2oj1e6cnr">Week 3 (6/15 - 6/19)</a>
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | <a href="https://docs.google.com/document/d/1HvOEUNTXplBFVLyiLFqUM917iPTD_HMbZdsruq0erJE/edit#heading=h.xgegbjwvfyf">Week 4 (6/22 - 6/26)</a>
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | <a href="https://docs.google.com/document/d/1HvOEUNTXplBFVLyiLFqUM917iPTD_HMbZdsruq0erJE/edit#heading=h.3ael4skbu2k4">Week 5 (6/29 - 7/3)</a>
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | <a href="https://docs.google.com/document/d/1HvOEUNTXplBFVLyiLFqUM917iPTD_HMbZdsruq0erJE/edit#heading=h.gvbqdzb1kqf6">Week 6 (7/6 - 7/10)</a>
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | <a href="https://docs.google.com/document/d/1HvOEUNTXplBFVLyiLFqUM917iPTD_HMbZdsruq0erJE/edit#heading=h.ebadnxicnqsi">Week 7 (7/13 - 7/17)</a>
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | <a href="https://docs.google.com/document/d/1HvOEUNTXplBFVLyiLFqUM917iPTD_HMbZdsruq0erJE/edit#heading=h.qjnk87xe52sy">Week 8 (7/20 - 7/24)</a>
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | <a href="https://docs.google.com/document/d/1HvOEUNTXplBFVLyiLFqUM917iPTD_HMbZdsruq0erJE/edit#heading=h.9ytnncddh38t">Week 9 (7/27 - 7/31)</a>
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | <a href="https://docs.google.com/document/d/1HvOEUNTXplBFVLyiLFqUM917iPTD_HMbZdsruq0erJE/edit#heading=h.v1nevkzea28q">Week 10 (8/3 - 8/7)</a>
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | <a href="https://docs.google.com/document/d/1HvOEUNTXplBFVLyiLFqUM917iPTD_HMbZdsruq0erJE/edit#heading=h.x44heahdq0mm">-Skip Week 11-</a>
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | <a href="https://docs.google.com/document/d/1HvOEUNTXplBFVLyiLFqUM917iPTD_HMbZdsruq0erJE/edit#heading=h.nm5utcm00u80">Week 12 (8/17 - 8/21)</a>
| + | |
| − |
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <br/>
| + | |
| − | <br/>
| + | |
| − | <br/>
| + | |
| − | <br/>
| + | |
| − | <br/>
| + | |
| − | <br/>
| + | |
| − | <br/>
| + | |
| − | <br/>
| + | |
| − | <br/>
| + | |
| − | <br/>
| + | |
| − | <br/>
| + | |
| − | <br/>
| + | |
| − | <br/>
| + | |
| − | <br/>
| + | |
| − | <br/>
| + | |
| − | <br/>
| + | |
| − | <br/>
| + | |
| − | <br/>
| + | |
| − | <br/>
| + | |
| − | <br/>
| + | |
| − | <br/>
| + | |
| − | <br/>
| + | |
| − | <br/>
| + | |
| − | <br/>
| + | |
| − | <h1 dir="ltr">
| + | |
| − | Week 2 (6/8 - 6/12)
| + | |
| − | </h1>
| + | |
| − | <p dir="ltr">
| + | |
| − | 6/8/2015
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for the day:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run a gel after digestion of lacZ and pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Gel extraction & digest cleanup
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Ligation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transformation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Ran a gel after digestion
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Performed gel extraction and digestion cleanup
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Measured concentrations of pDawn and lacZ with nanodrop
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | PCR clean-up for new PCR of lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digestion of lacZ and pDawn with new Nde1 and BamHI
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digest clean-up
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Ligation of lacZ and pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux box transformation into E. coli
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 6/9/2015
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | PCR cleanup for lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digest lacZ and pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digest cleanup
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Ligation of pDawn and lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux box transformation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Design one plasmid system with Lux box/lacZ and design two plasmid system, one with Lux box and the other with the lacZ reporter
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | PCR cleanup for lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digest of lacZ and pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digest cleanup of lacZ and pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Ligation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux box transformation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Began working on plasmid design
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculated pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Potentially design primers for cloning Lux box into pDawn/lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Design SapI primers
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Design compatibility system
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pick colonies for BioBrick, inoculate, glycerol stock
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Colony PCR pDawn + lacZ, glycerol stock, miniprep, sequencing
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Induce lux box w/ arabinose
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Questions:
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | In the “empty” region of pDawn (the only space we think a biobrick can be cloned) there’s only
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | one restriction enzyme that cuts once in that region and nowhere else, SapI.
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Are we right in assuming that empty space is the only place we can clone in a biobrick?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Would it be okay if we used only one restriction enzyme for the ligation instead of two?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 6/10/2015
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Design SapI primers
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Design Compatibility Systems
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pick Colonies for Biobrick, inoculate, glycerol stock
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Colony PCR, pDawn + LacZ, Glycerol stock, miniprep
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Redigest pDawn (with extreme controls)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Reattempt Ligation of pDawn+LacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Colony PCR of BioBrick Transformation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run gel of colony PCR products
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run gel of newly digested pDawn, Overnight ligation.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Re-digest of pDawn (with extreme controls)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Re-attempt Ligation of pDawn+LacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Colony PCR of pDawn + LacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Gel extract of newly digested pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transformed lux box BioBrick, pDawn + lacZ, just pDawn backbone (for miniprep and
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | making our own glycerol stock) into NEB turbo again [had just cut pDawn backbone and just digested lacZ insert transformed as well as controls]
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plated transformed colonies and placed into incubator (will look at results tomorrow)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run gels of colony PCR from 6/10/15 and see if the lacZ was ligated into pDawn backbone
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish designing cloning of luxbox biobrick into pACBB-eGFP (plasmid that is compatible with pDawn)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pick colonies for most recent BioBrick Transformation, Inoculate, glycerol stock, miniprep
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Colony PCR of new pDawn + LacZ transformation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Induce luxbox transformed E. coli with arabinose (growing up transformed E. coli to saturation, back diluting 1:100, growing up to OD600 of 0.3,
| + | |
| − | then inducing with low to high arabinose concentrations)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | If transformation results not looking good, we will need to re-digest lacZ insert and pDawn, and re-ligate pDawn into lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Questions:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | How exactly does the two-plasmid transformation work? Do we transform the NEB turbo with one and then make it competent again and transform it with
| + | |
| − | the second?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Is there any way we can check for incomplete digestion (because the gel resolution doesn't allow us to see that, since the part we cut out is only
| + | |
| − | a few bases big)?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 6/11/2015
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculate pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run gel of colony PCR
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Check plates from yesterday
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Colony PCR of re-transformed plates (after we miscalculated the ligation of the first plates)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digest and ligate pACBB-eGFP and lux box
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up luxbox
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculate pACBB-eGFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculated pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Ran gel of first colony PCR
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Checked plates grown overnight
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Performed second colony PCR of newly transformed plates with pDawn + lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grew up luxbox transformants in LB+chlor (see double check in transformation worked because Chlor diffused in plate)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculated DH5alpha with pACBB-eGFP (in prep for miniprep and glycerol stock tomorrow)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Retransformed lux box biobrick into NEB turbo
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep and glycerol stock pACBB-eGFP inoculants
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Check inoculated colonies from luxbox transformation attempt #2 to see if any grew in LB+Chlor
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Look at transformation results on plates (for attempt #3 of transforming lux box biobrick)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Look at results for colony PCR of pDawn + lacZ (run gel + image)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Questions:
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | None, we might have some after we get the colony PCR results.
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 6/12/2015
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep pACBB-eGFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock of pACBB-eGFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Back dilute/ induce lux box→ Back dilute @ 12:57 PM
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Colony PCR lux box→ run on a gel
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock lux box
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock of pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 3 hr digest of pDawn + lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Overnight ligation of pDawn + lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run colony PCR on a gel for pDawn + lacZ ligation product.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pCR lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep pACBB-eGFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock of pACBB-eGFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Colony PCR lux box→ run on a gel
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock of pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run colony PCR on a gel for pDawn + lacZ ligation product.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Monday:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Induce lux box
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock lux box
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 3 hr digest of pDawn + lacZ (didn’t do before because needed more lacZ)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Overnight ligation of pDawn + lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Clone lux box into pACBB-eGFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Questions:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Does completing PCR on LacZ as a positive control using the Colony PCR Protocol (a.k.a using Taq instead of Phusion) have a significant effect on
| + | |
| − | the results of the PCR? [If not clear, this question is better explained in person]
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Should we purchase the enzymes and dntps? How long after the expiration date are the enzymes good for? Should we get rid of enzymes/ dntps that are
| + | |
| − | months/years past their expiration date?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | What happens if you use a buffer for restriction digest that is not the optimal one?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <h1 dir="ltr">
| + | |
| − | Week 3 (6/15 - 6/19)
| + | |
| − | </h1>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | -6/15/2015 SKIPPED-
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | 6/16/2015
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for today:
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Innoculate pACBB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make M9 media
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Perform gel extraction on pDawn, lacZ, pACBB-eGFP, and luxbox from gel we ran yesterday.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Ligate luxbox and pACBB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | LIgate pDawn and lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform luxbox and pACBB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform pDawn and lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Back dilute and induce luxbox
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Measure luminescence of induced luxbox on the hour
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | PCR cleanup on lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 3 hour digest of lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Made M9
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Gel extracted pDawn, lacZ, luxbox, and pACBB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Ligated luxbox and pACBB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Ligated pDawn and lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transformed luxbox and pACBB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transformed pDawn and lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Back diluted and induced luxbox
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Measured luminescence of induced luxbox on the hour
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | PCR clean up on lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digested lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Innoculate pACBB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for tomorrow:
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock luxbox and pSB1C3
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep more pACBB-eGFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Look at transformed plates, pick colonies, innoculate
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Colony PCR lacZ from pDawn and luxbox from pACBB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Column purify digested lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Dilute luxbox to OD600 .3, take measurements at 1 hr, 3 hr, 5 hr, etc.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Take 24th hour time point for lux box trial 1 (the one we started today)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Potentially do overnight ligations if the transformations from today don’t work
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep and glycerol stock ligated plasmids pDawn and lacZ and luxbox and pACBB.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 6/17/2015
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock LuxBox + PSB1C3
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep more pACBB-eGFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Look at plates, pick colonies, inoculate at 4pm LuxBox+pACBB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Colony PCR luxbox from pACBB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Column purify digested lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform overnight ligation (& prep plates)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | miniprep and glycerol stock ligated pDawn+LacZ and Luxbox+pACBB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform BL21 with luxbox biobrick.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock of LuxBox+PSB1C3
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep more pACBB-eGFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Design primer for lacZ RE site extensions
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Design luxbox colony PCR primers
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Design luxbox sequencing primers
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Design lacZ sequencing primers
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Look at plates (ligation products lacZ + pDawn and lux + pACBB), pick colonies, innoculate.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plot data of lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculate BL21 w/ lux biobrick (to back dilute on Fri)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Friday:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep + glycerol stock lacZ + pDawn and lux + pACBB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Dilute BL21 w/ lux biobrick to OD600 .3
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Take measurements at 1 hr, 3 hr, 5 hr, etc.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plot data ^
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Questions:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Is N = 2 enough for NEB Turbo for measuring luminescence experiment (as opposed to BL21 which we will do later)? Should we do more trials?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Should we do the interlab study this year? ( <a href="https://2015.igem.org/Tracks/Measurement/Interlab_study">https://2015.igem.org/Tracks/Measurement/Interlab_study</a>)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | We should discuss human practices
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 6/18/2015
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculate BL21 LuxBox + PSB1C3
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Colony PCR of pDawn + LacZ transformation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculate pACBB + LuxBox, pDawn + LacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform pDawn + LacZ from Mike into Turbo Cells
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digest test for pDawn + LacZ; pACBB + Lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol Stock BL21 with LuxBox
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order LacZ Assay Kit
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order pDawn & pACBB insert primers
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order Lux Sequence primers.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make more arabinose, autoclave flasks, prep for BL21 vs. Turbo experiment.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make LB Chlor. plates
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculate BL21 LuxBox + PSB1C3
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculate pACBB + LuxBox, pDawn + LacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform pDawn + LacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order pDawn & pACBB insert primers
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Find LacZ Assay kit we want to order (See links below)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order Lux Sequence primers.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make more arabinose, autoclave flasks, prep for BL21 vs. Turbo experiment.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol Stock BL21 + LuxBox
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digest Test of pDawn + LacZ & pACBB + LuxBox
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave LB Agar & pour plates ( Add Chlor)!!
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Back dilute & induce BL21 and NEB Lux strains
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Take luminescence measurements.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 6/19/2015
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol Stock BL21 + LuxBox
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digest Test of pDawn + LacZ & pACBB + LuxBox
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave LB Agar & pour plates
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pick colonies pDawn + lacZ; inoculate
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock pDawn + lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock 6 colonies for BL21 vs turbo experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol Stock BL21 + LuxBox
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digest Test of pDawn + LacZ & pACBB + LuxBox
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave LB Agar & pour plates
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pick colonies pDawn + lacZ; inoculate
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock pDawn + lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock 6 colonies for BL21 vs turbo experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Monday:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Back dilute & induce BL21 and NEB Lux strains
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Take luminescence measurements.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculate 10 ml cultures
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <h1 dir="ltr">
| + | |
| − | Week 4 (6/22 - 6/26)
| + | |
| − | </h1>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Back-dilute to 0.3
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Measure luminescence at 1h, 3h, etc.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Presentation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | lacZ expression as a result of light
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB vs. PSBIC3
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | BL21 vs. turbo in lux exp.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | lux exp. high vs low AHL →
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | questions
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | GANT for summer
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 20 kb from IDT
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | nonhydrolyzable arabinose
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Josh + Danny at 1 PM
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Decide if digest test worked
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol start="6">
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Back-dilute to 0.3
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Measure luminescence at 1h, 3h, etc.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Presentation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | lacZ expression as a result of light
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB vs. PSBIC3
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | BL21 vs. turbo in lux exp.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | lux exp. high vs low AHL →
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | questions
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | GANT for summer
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 20 kb from IDT
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | nonhydrolyzable arabinose
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Josh + Danny at 1 PM
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Decide if digest test worked
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Monday:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze luxbox in BL21 versus NEB Turbo
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform ligated pDawn+lacZ and pACBB+lux, then induce with arabinose
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Explore IDT 20kb options
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Figure out our goals (given our knowledge of Peking)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Work on website design
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Work on human practices
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 6/23/15
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims For Today:
| + | |
| − | </p>
| + | |
| − | <ol start="7">
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze luxbox in BL21 versus NEB Turbo
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform ligated pDawn+lacZ and pACBB+lux, then induce with arabinose
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Explore IDT 20kb options
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Figure out our goals (given our knowledge of Peking)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Work on website design
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Work on human practices
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplished Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyzed luxbox in BL21 versus NEB Turbo data
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transformed pDawn+lacZ and pACBB+lux into BL21
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Looked into other past iGEM team projects involving communication through light
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Started thinking about human practices and figuring out the logistics of sorting past projects by topic/team/year
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims For Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Compare fluorescent output for BL21 at different ODs by taking measurements 1hr, 3hr, 5hr, etc.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Questions/Concerns:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Discuss what the other teams are doing/have done and how are goals should shift accordingly
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 6/24/15
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims For Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Back dilute luxbox cultures to different OD’s, induce, measure OD until 7 hrs
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Decide on 20 kb from IDT
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Listen to IDT webinar @2 pm
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Look @ transformed plates and innoculate
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock ligated pDawn + lacZ and pACBB + lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Look over lacZ kit
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up cultures for both (ours and Mike’s) pDawn lacZ experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock Mike’s pDawn+lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up lux BL21 cultures for arabinose spiking experiments
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order d-fucose
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims For Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn+lacZ experiment (varying blue light intensity)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | back dilute lux BL21 to .3 and induce with arabinose
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Measure lux BL21 and add arabinose every hour
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pick up dNTPs
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 6/25/15
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims For Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Re-do yesterday’s experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pick up dNTPs
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | order more SOC
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | make color chart _ plan prospective experiments
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | IDT → $$
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | plan lacZ + pDawn experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Research previous team projects
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Created a spreadsheet detailing all sender-xreceiver cell relationships
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Rethought lux box experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Set up tomorrow’s pDawn + lacZ experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculated cultures for ^ experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Complete pDawn + lacZ experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pick up dNTPs
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order more SOC
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | IDT → $$
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Karol’s iGEM database project
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 6/26/15
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims For Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | iGEM database (human practices)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | B-Galactosidase assay
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finalize plan for the summer
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | OD + luminescence experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | iGEM database (human practices)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | B-Galactosidase assay
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | OD + luminescence experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Monday:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Co-transforming pDawn + lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Redoing standardized curve for B-Galactosidase assay
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Design exp for different blue light intensity on B- Galactosidase activity experiment.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + pSB1C3 experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Did pACBB + lux ligate properly? (we learned that pDawn + lacZ cloning did not work, but we still have Mike’s which turned yellow when we did
| + | |
| − | the assay yay!)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Decide which parts to order from IDT (but actually though)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Examine more of the different color systems that we have been discussing; talk to advisors about this more; decide which we want to do
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Questions:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | <a href="https://docs.google.com/spreadsheets/d/1Hos0nFdv6kmtn_Q1y98bH6RwkHVTM650sZQLPafrFRg/edit?usp=sharing">
| + | |
| − | https://docs.google.com/spreadsheets/d/1Hos0nFdv6kmtn_Q1y98bH6RwkHVTM650sZQLPafrFRg/edit?usp=sharing
| + | |
| − | </a>
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | (Just a quick conclusion) Finished the assay and noticed that only Mike’s pDawn + lacZ turned yellow after adding WS; our ligation was not
| + | |
| − | successful
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | But, when we tried to make the standard curve, we got a graph that didn’t make any sense.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | We think it could be because:
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | The equation includes OD660 for something that’s not a cell?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | We waited too long to add the stop solution (too yellow)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | We diluted many times to get the OD420 in the range of 0.2 - 1.0; we may have not mixed the solutions enough (the ODs were all pretty
| + | |
| − | much 0.6)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <h1 dir="ltr">
| + | |
| − | Week 5 (6/29 - 7/3)
| + | |
| − | </h1>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 6/29/15
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims For Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish presentation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | DIscuss project ideas; goals and data with advisors during Monday Meeting
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Figure out trajectory for the summer.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish presentation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | DIscuss project ideas; goals and data with advisors during Monday Meeting
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Figure out trajectory for the summer.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Co transform pDawn + lacZ + luxbox (fail-fast)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Redo standard curve for B- Gal assay
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Design experiment for blue-light intensity for assay
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Did lux box + pACBB ligate properly?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Decide which parts to order from IDT (but actually)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Talk about different color circuits
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 6/30/15
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims For Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | grow up pACBB-lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | dilute B-Gal
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up BL21 lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up lux for spiked arabinose experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Fill out safety form
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Set up incubator for blue light vs. dark pDawn experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Get primers
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Sequence pACBB-lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculate pDawn-lacZ for blue light inducibility experiment (dark)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Set up sequencing tubes for pACBB-lux part
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | do colony PCR at pACBB-lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | redo B-gal assay standardized curve
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up BL21 lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up pACBB-lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up lux for spiked arabinose experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Fill out safety form
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Set up incubator for the blue light vs. dark pDawn experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Get primers
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Set up sequencing tubes for pACBB-lux part
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | back dilute pDawn - lacZ 1:100 (dark)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | at OD of around .4, separate→ 3 blue; 3 dark
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | miniprep pACBB lux (2 tubes)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Sequence miniprepped pACBB-lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | PCR test of miniprepped pACBB-lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Redo B-gal assay standardized curve
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Redesign the illuminated incubator
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Friday:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | After 20 h of growth, do OD600 and miller assay for pDawn lacZ light vs. dark
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/1/15
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave flasks
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Set up blue light incubator
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At OD of .4, induce pDawn-lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Back dilute pDawn-lacZ 1:100 (dark) in 25 mL LB Kan
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Dilute B-Gal for curve
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Do B-Gal curve w/ BL21
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep pACBB-lux (4 tubes)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Back-dilute 1:100 lux BL21 for arabinose experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Induce lux w/ arabinose every hour; measure OD + luminescence
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave flasks
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Colony PCR
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Sequencing.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave flasks
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Set up blue light incubator
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At OD of .4, induce pDawn-lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Back dilute pDawn-lacZ 1:100 (dark) in 25 mL LB Kan
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Dilute B-Gal for curve
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Do B-Gal curve w/ BL21
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep pACBB-lux (4 tubes)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Back-dilute 1:100 lux BL21 for arabinose experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Induce lux w/ arabinose every hour; measure OD + luminescence
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave flasks
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Colony PCR
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run PCR of pACBB lux on gel
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pour gel
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Send in sequencing of pACBB-lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At ll AM, miller assay of pDawn light vs. dark
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | If pACBB gel good, do pACBB vs. pSB1C3 experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/2/15
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Sequencing of pACBB-lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pour gel
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run pACBB on gel
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At 11 AM, Miller Assay of pDawn light vs. dark
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Co-culture pSB1C3 + lux and pDawn + lacZ. You have to cotransform the opposite antibiotic resistance into each plasmid before transformation.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Dialysis Tube (ask Mike for this) _ controls for them growing at different rates
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Just pipet them into the same tube
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up a giant culture of pSB1C3 + lux; induce at the right times for visible luminescence
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | reach out to the iGEM teams who did similar experiments to see what they did for a visible glow.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Sequencing of pACBB-lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pour gel
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run pACBB on gel
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At 11 AM, Miller Assay of pDawn light vs. dark
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run PCR of pACBB-lux on gel
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pour gel
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Send in sequencing of pACBB-lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At ll AM, miller assay of pDawn light vs. dark
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | If pACBB gel good, do pACBB vs. pSB1C3 experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <h1 dir="ltr">
| + | |
| − | Week 6 (7/6 - 7/10)
| + | |
| − | </h1>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/6/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave flasks
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miller assay of samples from light vs. dark experiment (samples were stored in the fridge for 4 days, so if it doesn’t work, restart experiment)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Send in sequencing
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 pDawn-lacZ in 25 mL LB-Kan
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Redo PCR of lux-pACBB; troubleshoot?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculate pACBB-lux from glycerol stock
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At an OD of 0.3, induce with arabinose measure luminescence every hour (controls: uninduced pACBB-lux and pACBB-eGFP)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At an OD of 0.3, induce with blue light
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze arabinose spiking data.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave flasks
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miller assay of samples from light vs. dark experiment attempted_ experiment restarted
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Send in sequencing
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 pDawn-lacZ in 25 mL LB-Kan
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Redo PCR of lux-pACBB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculate pACBB-lux from glycerol stock
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At an OD of 0.3, induce with arabinose measure luminescence every hour (controls: uninduced pACBB-lux and pACBB-eGFP)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At an OD of 0.3, induce with blue light
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze arabinose spiking data.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | > Debrief from monday meeting
| + | |
| − | </p>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | sequence Mike’s pDawn+lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | research lacZ degradation rate (decide if we need to characterize)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | if we can’t find much, expose pDawn lacZ to blue light for 2h then measure lacZ expression every 2 hours until 20 h
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | clone pDawn essentials onto pACBB backbone (make primers to PCR off pDawn portion needed)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | adjust beta-galactosidase curve for concentration
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | discuss Karol database stuff (we disagree with Monday’s discussion)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | be more careful with what we put in the Monday meeting slide
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | don’t put in tentative data
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | keep it short
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | focus on the questions you have
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | focus on graphs, experimental designs, conclusions
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | data into excel spreadsheet
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | talk to Victor about lacZ, decide whether it’s worth it to maybe switch to GFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | ask Victor about how quickly it degrades
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | as soon as sequencing results come back, cotransform
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | either with pACBB or other compatible plasmids with the opposite antibiotic resistance
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | do an experiment with
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | do the poster 2 weeks before the July 31st deadline
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | present it at the meeting, so we can nip any potential problems in the butt
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | talk about restructuring our day and integrating most poster/website stuff
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | include laying out experimental designs and sending them out
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | </ul>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Questions:
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/7/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miller assay
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Arabinose spiking experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Research on lacZ degradation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pour gel
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up from glycerol stock *B2* BL21 lux for OD experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run gel of pACBB PCR
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Talk to Seville about equipment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Sequence pDawn+lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plan lacZ experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Look into XGal
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow Mike’s pDawn *get from Mike
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Figure out luminometer
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up pDawn-LacZ (A1)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miller assay
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pour gel
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run gel of pACBB PCR
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plan lacZ-degrading experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 for OD experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Induce with arabinose at dif OD and take luminescence measurements
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Illuminometer meeting with Mike
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 of pDawn-lacZ in 25 mL
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At OD of 0.4, induce with blue light
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Measure lacZ every 3 hours
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/8/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Sequence rest of pACBB-lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Sequence pDawn-lacZ (Mike’s) with primers that bind to outside
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pour gel
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run gel of pDawn+lacZ PCR
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Do pDawn+mRFP light vs. dark experiment (1:100 at 11 AM and induce at 2 PM)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 BL21 lux in 25 mL LB+chlorom
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Design primers to swap out promoter in lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB vs. pSBIC3 experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Backdiluted BL21 and DH5-alpha with pDawn RFP 1:100 for blue vs. dark experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Induced pDawn RFP strains at OD ~.3-.4 with blue light
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Poured gel, ran gel of pDawn lacZ PCR check → success! lacZ amplified, next step if to sequence pDawn lacZ to make sure the band is what we think
| + | |
| − | it is
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculated BL21 lux from glycerol stock for arabinose induction at different OD experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Designed primers to change lux operon promoter to T7 (not pBAD) by cloning into pRSFDuet and pET-26b(+) backbones
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Backdiluted pACBB lux 1:100, induced with arabinose at OD ~.4, took OD measurements every hour for 7h (to characterize pACBB lux expression vs.
| + | |
| − | pSCB1C3 lux expression)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Backdilute BL21 lux 1:100 in LB Chlor (9 AM)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Measure RFP expression in pDawn blue light vs. dark experiment at 20 h (10 AM)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Backdilute pDawn lacZ 1:100 in 25 mL LB Kan to redo blue vs. dark experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | induce pDawn lacZ with blue light at OD ~.4
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep pDawn lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Send of pDawn lacZ for sequencing with primers (2) that bind to outside pDawn
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze pACBB vs. pSB1C3 luminescence results
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Think about swapping out pDawn backbone for pACBB and redoing lacZ cloning (decide what to do moving forward)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Questions:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Should we move forward with changing the lux operon promoter to T7 ASAP, and should we order the primers for the cloning tomorrow?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/9/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock pDawn RFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Induce pDawn lacZ at OD .4 for light vs dark experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprepped pDawn RFP and pACBB lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Induce BL21 with arabinose at different OD and measured luminescence
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Sequence miniprepped pDawn lacZ with pDawn binding primers
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Sequence miniprepped pACBB lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze pACBB vs. PSB1C3 lux results
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Think about swapping out pDawn backbone
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze pDawn RFP light vs. dark results
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Cotransformed pDawn RFP and pACBB lux into BL21
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock pDawn RFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprepped pDawn RFP and pACBB lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyzed pACBB vs. PSB1C3 lux results
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyzed pDawn RFP blue light vs. dark results
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Looked into h-ns mutant E. coli and effects on lux operon expression
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Looked into other plasmids that would luminesce better than the lux operon biobrick
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Backdilute BL21 lux 1:100 in 40 mL LB Chlor for OD arabinose induction experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Sequence miniprepped pDawn lacZ with pDawn binding primers
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Sequence miniprepped pACBB lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Think about swapping out pDawn backbone
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Induce BL21 with arabinose at different OD and measured luminescence (started today but issues with Tecan readings)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pick colonies from transformation plates, inoculate, glycerol stock (glycerol stock later in day)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Look into ordering h-ns mutant E. coli and ordering plasmids off Addgene that may luminesce brighter than lux operon BioBrick
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Questions:
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/10/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Back dilute BL21 lux 1:100 for temperature experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Send off pDawn lacZ, pDawn RFP, and pACBB lux for sequencing
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | induce BL21 with arabinose and grow at different temperature
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pick colonies from cotransformation of pACBB lux and pDawn RPF
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Set up experiment to see if pACBB lux induced RPF expression
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | organize data
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | prepare BL21 competent cells
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | glycerol stock pDawn lacZ/RFP + pACBB lux BL21 strains from cotransformation (4 colonies per cotransformation)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Back dilute BL21 lux 1:100 for temperature experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Send off pDawn lacZ, pDawn RFP, and pACBB lux for sequencing
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Induce BL21 with arabinose and grow at different temperature, measure every hour
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pick colonies from cotransformation of pACBB lux and pDawn RPF
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Set up experiment to see if pACBB lux induced RPF expression
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finalize what to spend the 20kb on from IDT
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order primers for changing lux operon promoter
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculate pDawn RFP + pACBB lux colonies for fail fast attempt number two, backdilute 1:100 after grown to saturation, induce with arabinose at OD
| + | |
| − | ~.4
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | After 20h, measure RFP expression for all fail fast tubes (Tuesday)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze lux and pDawn sequencing results
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Email asking for the split lux plasmid from this (
| + | |
| − | <a href="http://link.springer.com/article/10.1007/s00216-010-4266-7/fulltext.html">
| + | |
| − | http://link.springer.com/article/10.1007/s00216-010-4266-7/fulltext.html
| + | |
| − | </a>
| + | |
| − | ); specifically asking for Sy102 and SY104
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Cotransform opposite antibiotic resistence for coculturing experiment (pUC57 Kan, find something for pDawn RFP)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Prepare BL21 competent cells (grow up cells in the morning to OD .3-.4 is the first step)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Questions:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | How do you make BL21 competent cells?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | What white microwell plates should we order?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Can we order primers to move ahead with replacing the pBAD promoter in lux with a T7 promoter?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | How should we plan the fail fast experiment? Right now, we have it set up so each cotransformed colony will grow in 1 tube with light (not induced
| + | |
| − | with arabinose to check for pDawn presence/activity alone) and two tubes in the dark (one induced with arabinose as our “test” tubes and one
| + | |
| − | uninduced to show that pDawn is not induced without lux operon expression). All of these are done in 1:1 LB kan LB Chlor media. We also have a
| + | |
| − | negative control of BL21 with the lux operon alone growing in LB kan+chlor to show that it will not grow. This tube will remain in the light and
| + | |
| − | idk if we should induce it with arabinose or not. Anyway, are there any other controls we should seriously consider including?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <h1 dir="ltr">
| + | |
| − | Week 7 (7/13 - 7/17)
| + | |
| − | </h1>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/13/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plan RFP-lux experiment #2
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Innoculate from glycerol stock
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plan Tecan calibration experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Ask Mike + Spencer to grant us access to BE undergrad lab
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze sequencing for pDawn lacZ and lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Innoculate from glycerol stock
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plan Tecan calibration experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Prepare presentation.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/14/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave flasks
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make BL21 competent cells (6; grow up 1)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make LB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make LB + chlor
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Co transform w/ opposite antibiotic
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Ask Mike for pet26B
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order primers to swap out GFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Tecan test
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | inoculate A5 and B2/B3
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Print a toucan
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 pDawn + lacZ; At an OD of 0.4 induce
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 cotransformed stuff; At an OD of 0.4 induce
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make arabinose
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order luminol + email iGEM team
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze sequencing results.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave flasks
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze sequencing results.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make arabinose
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 cotransformed stuff; At an OD of 0.4 induce
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculate
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Tecan test-
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | A1 - Mike’s ligated pDawn + lacZ (as a negative control)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | B2 - pSB1C3 + lux box (to test luminescence)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miller Assay (light vs. dark)-
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | A1 - Mike’s ligated pDawn + lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | B2 - pSB1C3 + lux box (negative control)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | BL21 + lux - USE B2
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Mike’s ligated pDawn +lacZ - USE A1
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux - USE A7
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pick up LB from the autoclave
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make BL21 competent cells
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make BL21 glycerol stock
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Tecan test to make sure that luminescence measurements are correct
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Co transform w/ opposite antibiotic
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Ask Mike for pet26B
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order primers to swap out GFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 pDawn + lacZ.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Induce with blue light at an OD of 0.4
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculate cotransformed pACBB + lux box & pDawn + lacZ for experiment tomorrow.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze data for cotransformed mRFP experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | START POSTER
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Thursday:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run two experiments simultaneously
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Nikita/Hannah
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miller assay for pDawn + lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | After 20 hours, measure lacZ expression
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze data
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Jane/Karol
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Repeat cotransformed mRFP experiment with cotransformed lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Based on results from the first experiment, validate the second experiment.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/15/15
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make new LB (since we messed it up)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make BL21 competent cells
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make BL21 glycerol stock
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up pet26B
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 pDawn + lacZ.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Induce with blue light at an OD of 0.4
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculate cotransformed pACBB + lux box & pDawn + lacZ for experiment tomorrow.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze data for cotransformed mRFP experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make new LB (since we messed it up) + LB chlor
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up pet26B
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 pDawn + lacZ.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Induce with blue light at an OD of 0.4
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inoculate cotransformed pACBB + lux box & pDawn + lacZ for experiment tomorrow.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze data for cotransformed mRFP experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pla big flask culture experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 20 hr of pDawn-lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 20 hr of RFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run big flask culture experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze RFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn-LacZ Failfast Experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At 9AM do a 1:100 dilution
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Set up PET primers
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Design high GC-lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Email Cambridge iGEM
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Black well plates
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order luminol
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order primers to swap out GFP and RFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/16/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 20 hr of pDawn-lacZ
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 20 hr of RFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run big flask culture experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze RFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn-LacZ Failfast Experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At 9AM do a 1:100 dilution
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Set up PET primers
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Design high GC-lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Email Cambridge iGEM
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Black well plates
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order luminol
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order primers to swap out GFP and RFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 20 hr of pDawn-lacZ light vs. dark experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 20 hr of RFP reading
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Run big flask culture experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze RFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn-LacZ Failfast Experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At 9AM do a 1:100 dilution
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miller assay at 3 time points (2,5 and 7 hours)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | OD and luminescence readings every hour
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Set up PET primers
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Email Cambridge iGEM
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order black well plates
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze data from pDawn-lacZ failfast experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze data from pDawn-RFP failfast experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Determine if the failfast worked
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Start the Monday Meeting presentation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Talk about final goals
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/17/15
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze data from pDawn-lacZ failfast experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze data from pDawn-RFP failfast experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Determine if the failfast worked
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Start the Monday Meeting presentation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Talk about final goals
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze data from pDawn-lacZ failfast experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze data from pDawn-RFP failfast experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Determine if the failfast worked
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Start the Monday Meeting presentation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Talk about final goals
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Monday:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Determine direction
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish Monday Meeting presentation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <h1 dir="ltr">
| + | |
| − | Week 8 (7/20 - 7/24)
| + | |
| − | </h1>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/20/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today/Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish analyzing data
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Prepare for Monday Meeting presentation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Talk more about our goals and direction now that the failfast didn’t “work”
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | How do we get it to glow?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform pACBB+lux and pSB1C3 + lux in Top10/ NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make the photon counter <- HIGHEST PRIORITY
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order the inverter
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order T7 promoters
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Email Goulian about H-NS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Email about split lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Call Glowell
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order high GC lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/21/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform pACBB+lux and pSB1C3 + lux in Top10/ NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make the photon counter
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order the inverter
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order T7 promoters
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Email Goulian about H-NS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Email about split lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Call Glowell
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order high GC lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make the photon counter
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order T7 promoters
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Email about split lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Call Glowell
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order high GC lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform pACBB+lux and pSB1C3 + lux in Top10/ NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish the photon counter
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Order the inverter
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Email Goulian about H-NS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/22/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish the photon counter
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Email Goulian about H-NS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Poster deadline is approaching
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Work on the presentation for Seville’s talk
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make the photon counter
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Heard back about split lux: going to send it in two weeks if we want it
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Emailed Goulian and the lab Cambridge got their hns mutant strains from
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform into Top10/ NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 OF NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow to an OD of 0.4
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Spin them down
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pour out the LB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Resuspend in TSS buffer ← look in the Chow lab
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | NEB Turbo (- control)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | B9, B1, B4, G2 and cotransform G2 and F4
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Next week
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Leave plates out on the counter? On Monday, pick from them and ultimately make glycerol stock
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Redo lux experiments
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/22/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol start="6">
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish the photon counter
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Email Goulian about H-NS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Poster deadline is approaching
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Work on the presentation for Seville’s talk
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol start="4">
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make the photon counter
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Heard back about split lux: going to send it in two weeks if we want it
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Emailed Goulian and the lab Cambridge got their hns mutant strains from
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol start="2">
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform into Top10/ NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 OF NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow to an OD of 0.4
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Spin them down
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pour out the LB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Resuspend in TSS buffer ← look in the Chow lab
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | NEB Turbo (- control)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | B9, B1, B4, G2 and cotransform G2 and F4
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Next week
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Leave plates out on the counter? On Monday, pick from them and ultimately make glycerol stock
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Redo lux experiments
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/23/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | -Skip-
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/24/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make NEB10 competent
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 OF NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow to an OD of 0.4
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Spin them down
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pour out the LB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Resuspend in TSS buffer ← look in the Chow lab
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish working on the photon counter experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pick up primers from cell center
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish sending emails to source parts
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make NEB10 competent
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 OF NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow to an OD of 0.4
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Spin them down
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pour out the LB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Resuspend in TSS buffer ← look in the Chow lab
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish working on the photon counter experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol start="3">
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform into Top10/ NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | NEB Turbo (- control)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | B9, B1, B4, G2 and cotransform G2 and F4
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pSB1C3 + lux (B9)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | miniprepped pSB1C3 + lux (B1)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux (B4)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn + RFP (G2)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <br/>
| + | |
| − | <ol start="2">
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Next week
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Leave plates out on the counter? On Monday, pick from them and ultimately make glycerol stock
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Redo lux experiments
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <h1 dir="ltr">
| + | |
| − | Week 9 (7/27 - 7/31)
| + | |
| − | </h1>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/27/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform parts into NEB10 Competent Cells
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | B9, B1, B4, G2 and cotransform G2 and F4
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | NEB Turbo (- control)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pSB1C3 + lux (B9)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | miniprepped pSB1C3 + lux (B1)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux (B4)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn + RFP (G2)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish Monday Meeting presentation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finalize information for the website/poster
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plan experiment to measure luminescence with NEB10 tomorrow.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | “Big-ass culture” -->3 g/ 20 mL arab
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish photon counter (determine fate?)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Call Tecan
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Ask Jane
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | T7_pickup cell center
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Codon optimized
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inverter strain
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Send Fedex for Split Lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol start="8">
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform parts into NEB10 Competent Cells
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | B9, B1, B4, G2 and cotransform G2 and F4
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | NEB Turbo (- control)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pSB1C3 + lux (B9)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | miniprepped pSB1C3 + lux (B1)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux (B4)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn + RFP (G2)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish Monday Meeting presentation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finalize information for the website/poster
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plan experiment to measure luminescence with NEB10 tomorrow.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | “Big-ass culture” -->3 g/ 20 mL arab
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Determine fate of photon counter
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Call Tecan
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Ask Jane
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | T7_pickup cell center
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Codon optimized
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inverter strain
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Emailed Yolanda about Fedex for Split Lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plan out entire summer
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plan future experiments
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/28/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Flesh out goals for toolbox “story”
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform BBa_K525998 T7 + RBS Biobrick into competent cells (lux gene will be transformed into it to make pSB1C3 + lux)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Innoculate cultures
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn + mRFP in NEB10 (F4 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in NEB10 (G2 dna) X 4 (three from same colony to run big ass culture) ::::: (the other is to eventually use for pCR of part)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pSB1C3 + lux in NEB10 (B2 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprepped pSB1C3 + lux in NEB10 (B9 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Cotransformed (pDawn + mFRP + pACBB + lux) in NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in BL21 (DONE)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Obtain HNS strain form Goulian
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Flesh out goals for toolbox “story”
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform BBa_K525998 T7 + RBS Biobrick into competent cells (lux gene will be transformed into it to make pSB1C3 + lux)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Innoculate cultures
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn + mRFP in NEB10 (F4 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in NEB10 (G2 dna) X 4 (three from same colony to run big ass culture) ::::: (the other is to eventually use for pCR of part)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pSB1C3 + lux in NEB10 (B2 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprepped pSB1C3 + lux in NEB10 (B9 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Cotransformed (pDawn + mFRP + pACBB + lux) in NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in BL21 (DONE)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Obtain HNS strain form Goulian
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Figure out how to make T7 + pACBB + lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in NEB10 big ass culture to test for visible luminescence
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn + mRFP in NEB10 (F4 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in NEB10 (G2 dna) X 4 (three from same colony to run big ass culture) ::::: (the other is to eventually use for pCR of part)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pSB1C3 + lux in NEB10 (B2 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprepped pSB1C3 + lux in NEB10 (B9 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Cotransformed (pDawn + mFRP + pACBB + lux) in NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pick
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | HNS that Goulian gave us
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | BBa_K525998 T7 + RBS Biobrick
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Research
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | If we can use the T7 promoter in the HNS strain
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | If the HNS strain mutant (for AraD) so that the arabinose is not eaten up
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Creation of T7 + pSB1C3 + lux system
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep pACBB + lux in BL21 to later be used for pCR
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep BBa_K525998 T7 + RBS Biobrick
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plan for digestion, ligation and transformation on Thursday
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Give presentation to PFP scholars
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up pACBB + lux and cotransformed pACBB + lux & pDawn + mRFP for fail fast experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | TOOLBOX STORY → bolded combinations are prepared, but not tested. Italicized combinations are completed; nothing has been tested with nonanol; yellow are
| + | |
| − | knocked out because T7 promoter does not function in NEB10
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Possible combinations: TOOLBOX 1 LUMINESCENCE
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + T7 + high pSB1C3 + BL21 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + T7 + high pSB1C3 + NEB10 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + T7 + high pSB1C3 + HNS (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + T7 + low pACBB + BL21 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + T7 + low pACBB + NEB10 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + T7 + low pACBB + HNS (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + pBAD + high pSB1C3 + BL21 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + pBAD + high pSB1C3 + NEB10 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + pBAD + high pSB1C3 + HNS (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + pBAD + low pACBB + BL21 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + pBAD + low pACBB + NEB10 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + pBAD + low pACBB + HNS (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Possible combinations: TOOLBOX 2 SENDER RECEIVER SYSTEM (Simultaneously done)
| + | |
| − | </p>
| + | |
| − | <ol start="4">
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + T7 + high pSB1C3 + BL21 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + T7 + high pSB1C3 + NEB10 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + T7 + high pSB1C3 + HNS (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <ol start="4">
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + T7 + low pACBB + BL21 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + T7 + low pACBB + NEB10 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + T7 + low pACBB + HNS (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <ol start="4">
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + pBAD + high pSB1C3 + BL21 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + pBAD + high pSB1C3 + NEB10 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + pBAD + high pSB1C3 + HNS (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <ol start="4">
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + pBAD + low pACBB + BL21 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + pBAD + low pACBB + NEB10 (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux operon + pBAD + low pACBB + HNS (test with/without nonanol)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | IF TIME, TOOLBOX 2
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux AB + CDE
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux AB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Lux CDE
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | HUMAN PRACTICES
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Two inverter strains
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/29/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Figure out how to make T7 + pACBB + lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in NEB10 big ass culture to test for visible luminescence
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn + mRFP in NEB10 (F4 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in NEB10 (G2 dna) X 4 (three from same colony to run big ass culture) ::::: (the other is to eventually use for pCR of part)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pSB1C3 + lux in NEB10 (B2 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprepped pSB1C3 + lux in NEB10 (B9 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Cotransformed (pDawn + mFRP + pACBB + lux) in NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pick
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | HNS that Goulian gave us
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | BBa_K525998 T7 + RBS Biobrick
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Research
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | If we can use the T7 promoter in the HNS strain
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | If the HNS strain mutant (for AraD) so that the arabinose is not eaten up
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Creation of T7 + pSB1C3 + lux system
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep pACBB + lux in BL21 to later be used for pCR
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep BBa_K525998 T7 + RBS Biobrick
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plan for digestion, ligation and transformation on Thursday
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Give presentation to PFP scholars
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up pACBB + lux and cotransformed pACBB + lux & pDawn + mRFP for fail fast experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Figure out how to make T7 + pACBB + lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plan for pACBB + lux in NEB10 big ass culture to test for visible luminescence (there weren’t enough flasks to do it yesterday)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave flasks
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in NEB10 (G2 dna) X 4 (three from same colony to run big ass culture) ::::: (the other is to eventually use for pCR of part)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pSB1C3 + lux in NEB10 (B2 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprepped pSB1C3 + lux in NEB10 (B9 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Cotransformed (pDawn + mFRP + pACBB + lux) in NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pick
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | HNS that Goulian gave us
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | BBa_K525998 T7 + RBS Biobrick
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn + mRFP in NEB10 (F4 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Research
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | If we can use the T7 promoter in the HNS strain
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | If the HNS strain mutant (for AraD) so that the arabinose is not eaten up
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Creation of T7 + pSB1C3 + lux system
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep pACBB + lux in BL21 to later be used for pCR
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plan for digestion, ligation and transformation on Thursday
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow::
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish the poster
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in NEB10 in big ass culture
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep BBa_K525998 T7 + RBS Biobrick
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 HNS at 11AM
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make HNS cells competent
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol Stock pDawn + mRFP in NEB10 (F4 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Cleanup pCR product (lux)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digestion of pCRed lux + miniprepped Biobrick
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform HNS with pBAD + pACBB + lux & pBAD + pSB1C3 + lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Overnight ligation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/30/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish the poster
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in NEB10 in big ass culture
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep BBa_K525998 T7 + RBS Biobrick
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Cleanup pCR product (lux)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digestion of pCRed lux + miniprepped Biobrick
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Overnight ligation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 HNS at 11AM
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make HNS cells competent
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up HNS for big culture tomorrow
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock HNS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform HNS with pBAD + pACBB + lux & pBAD + pSB1C3 + lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol Stock pDawn + mRFP in NEB10 (F4 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave BE lab flasks and flasks for big ass HNS culture tomorrow
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Bleach everything on the table in the back room
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Talk more about switching out the promoter; ask the advisors if we should bother choosing
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in NEB10 in big ass culture
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Miniprep BBa_K525998 T7 + RBS Biobrick
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Cleanup pCR product (lux)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digestion of pCRed lux + miniprepped Biobrick + cleanup
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 HNS at 11AM
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make HNS cells competent
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up HNS for big culture tomorrow
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock HNS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform HNS with pBAD + pACBB + lux & pBAD + pSB1C3 + lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol Stock pDawn + mRFP in NEB10 (F4 dna)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave BE lab flasks and flasks for big ass HNS culture tomorrow
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Bleach everything on the table in the back room
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Talk more about switching out the promoter; ask the advisors if we should bother choosing
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digestion + Cleanup
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Overnight ligation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Big ass culture for HNS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock HNS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform HNS with pBAD + pACBB + lux & pBAD + pSB1C3 + lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave BE lab flasks
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Talk more about what final goals will be
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish poster
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Figure out luminometer
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | If time, transform T7 promoter construct into BL21; also cotransform with pDawn + mRFP (GROW UP IN THE MORNING)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 cotransformed cultures
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Measure OD, luminescence and RFP every hour
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Questions:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Should we use the T7 promoter if we cannot put it into HNS or NEB10? Does it detract from the comprehensiveness of the toolbox?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 7/31/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digestion + Cleanup
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Overnight ligation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Big ass culture for HNS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock HNS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform HNS with pBAD + pACBB + lux & pBAD + pSB1C3 + lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave BE lab flasks
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Talk more about what final goals will be
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish poster
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Figure out luminometer
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | If time, transform T7 promoter construct into BL21; also cotransform with pDawn + mRFP (GROW UP IN THE MORNING)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 cotransformed cultures
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Measure OD, luminescence and RFP every hour
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Digestion + Cleanup
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock HNS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Transform HNS with pBAD + pACBB + lux, pBAD + pSB1C3 + lux, pBAD + pACBB, pDawn + mRFP and cotransformed pACBB + lux and pDawn + mRFP
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Autoclave BE lab flasks
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Complete poster (or just generate an idea to be completed on the weekend)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Monday:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Do the cotransformed mRFP experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 cotransformed cultures
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Measure OD, luminescence and RFP every hour
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Talk more about final goals
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Iron out the experiments that we have planned for the rest of the summer
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | tracking
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | proving orthogonality
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Prepare final poster presentation to send out to the advisors on Sunday night
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Prepare for Tuesday Advisor Meeting
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <h1 dir="ltr">
| + | |
| − | Week 10 (8/3 - 8/7)
| + | |
| − | </h1>
| + | |
| − | <p dir="ltr">
| + | |
| − | 8/3/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Do the cotransformed mRFP experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 cotransformed cultures
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Measure OD, luminescence and RFP every hour
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Talk more about final goals
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Iron out the experiments that we have planned for the rest of the summer
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | tracking
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | proving orthogonality
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Prepare final poster presentation to send out to the advisors on Sunday night
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Prepare for Tuesday Advisor Meeting
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Talk more about final goals
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Iron out the experiments that we have planned for the rest of the summer
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Prepare for Tuesday Advisor Meeting
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Right now
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Orthogonality→ proving for sure that light and chemicals are orthogonal to each other
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Tracking → chemical vs. light
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | This experiment will demonstrate hopefully that light tracks better than chemicals which would mean it has better spatiotemporal
| + | |
| − | control
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | These two principles can be used to build a major experiment (Pim’s idea)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | A bidirectional communication system in which one cell communicates with the second cell with chemicals and the same second cell
| + | |
| − | communicates with the first using light. This should create a “stable” system which oscillates around a line because the K2 (light
| + | |
| − | communication) is higher than K1 (chemical communication). It is a form of negative feedback.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make adjustments to the presentation for advisor meeting with Dr. Chow
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Iron out more of the cloning details for Pim’s bidirectional idea
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make changes based on advisor feedback
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 8/4/15
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make adjustments to the presentation for advisor meeting with Dr. Chow
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Iron out more of the cloning details for Pim’s bidirectional idea ← Too complicated; not necessarily based on an engineering principle; our
| + | |
| − | abstract talks about this bidirectional communication, but our experiments are not necessarily aligning with it.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Rework the presentation to fit a more probable system that we can use.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | A semi-completed poster
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | What we thought was bistable communication, but instead was just a system that would permanently exist in the second state after addition of
| + | |
| − | light
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Prepare a distance experiment in which you have six tubes and place tubes inside them so that you can run “co-tubing experiment”
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up NEB10 glycerol stock one in the dark (pDawn + mRFP) and one in the light (pACBB + lux)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 dilute into flasks after reaching saturation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Measure at 20 hours after separating into individual tubes
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Adjust experiments based on if you can get this to work
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Prepare a list of principles that we want to base our entire project on; come up with a list of experiments that complement this list
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Ideas discussed during the meeting:
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pim’s tracking idea in which we mutate the lov domain to create different tracking curves
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Possibly a distance idea→ further iron out the application of this type of a project
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 8/5/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Prepare a distance experiment in which you have six tubes and place tubes inside them so that you can run “co-tubing experiment”
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up NEB10 glycerol stock one in the dark (pDawn + mRFP) and one in the light (pACBB + lux)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 dilute into flasks after reaching saturation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Measure at 20 hours after separating into individual tubes
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Adjust experiments based on if you can get this to work
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Prepare a list of principles that we want to base our entire project on; come up with a list of experiments that complement this list
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Prepare a list of principles that we want to base our entire project on; come up with a list of experiments that complement this list
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Current ideas on the table
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Light compatible with strains or environment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Tuneable photoreceptors with the lov mutation
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Luminescence signal → less persistent signal
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Chemical and light → different rates of production; orthogonality
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Light driven spatiotemporal control of chemical communication
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Ironed out specific experiments for the above
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Prepared the “co-tubing” experiment to run tomorrow
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Go through the principles/project ideas with advisors and narrow down to the ones we want
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish the co-tubing experiments and find out which ones are feasible ^
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish cloning the T7 promoter
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Characterize individual sender + receiver parts
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Find out what the sender cell curve looks like in NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Find out if the receiver responds to the sender cell; simulate the light curve to test this???
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plan experiments for the chosen project idea more thoroughly
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Redo poster for next Monday Meeting with Dr. Chow
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 8/6/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish ranking the projects/principles
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Take flasks out of autoclave at 12 PM
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 dilute them
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Induce at OD of 0.3
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Take 20 hr measurement tomorrow
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish pACBB + lux in NEB10 luminescence curve
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Inverter
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Identify 3 with different strengths
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Email advisors about projects; remind them that the abstract is due tomorrow
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Email advisors about projects; remind them that the abstract is due tomorrow
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Finish cloning T7
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Retransform into HNS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | F4
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | B9
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | G2
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | F4 + G2
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <h1 dir="ltr">
| + | |
| − | -Skip Week 11-
| + | |
| − | </h1>
| + | |
| − | <h1 dir="ltr">
| + | |
| − | Week 12 (8/17 - 8/21)
| + | |
| − | </h1>
| + | |
| − | <p dir="ltr">
| + | |
| − | 8/17/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | inoculate pdawn rfp (dh5alpha and BL21) for receiver characterization
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | inoculate psb1c3 lux in NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | backdilute psb1c3 lux at 6 PM
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | induced lux psb1c3 with arabinose at 10 PM (when OD at ~.4)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | order LB plates
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | finish soldering circuit for pDawn receiver characterization
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | setup tubes for pDawn receiver characterization experiment (to start tomorrow) (program arduino to turn off blue light at certain times)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | plate agar stab with split lux operon
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | plate agar stabs with each antibiotic only to separate plasmids
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | transform psb1c3 lux operon into h-ns
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | regraph pelleting volume luminescence data with photons on x-axis
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Publication for reference on split lux: http://link.springer.com/article/10.1007/s00216-010-4266-7/fulltext.html
| + | |
| − | </p>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | inoculate pdawn rfp (dh5alpha and BL21) for receiver characterization
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | inoculate psb1c3 lux in NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | backdilute psb1c3 lux at 6 PM
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | induced lux psb1c3 with arabinose at 10 PM (when OD at ~.4)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | order LB plates
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | finish soldering circuit for pDawn receiver characterization
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | plate agar stab with split lux operon
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | plate agar stabs with each antibiotic only to separate plasmids
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | transform psb1c3 lux operon into h-ns
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tuesday:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | backdilute pDawn RFP 1:100 for receiver characterization
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | induce pDawn RFP with light at 10 AM (when OD at ~.4)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | turn off blue light at appropriate times for pdawn receiver characterization (or check that arduino is working)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 8 AM → take a 10 h timepoint for luminescence of psb1c3 lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | take hourly luminescence and OD readings for psb1c3 lux up to 17 h (3PM)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pick split lux operon colonies, inoculate (at least 3 colonies)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pick and replate colonies for attending to separate out plasmids
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pick h-ns lux transformed colonies, inoculate
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 8/18/15
| + | |
| − | </p>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Today:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | glycerol stock h-ns transformed with psb1c3 lux (at least 3 colonies)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | glycerol stock split lux operon colonies (at least 3 colonies)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | inoculate h-ns lux from glycerol stock for time course experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pick and replate colonies for attending to separate out plasmids for second time
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Replan NEB10 pSB1C3 + lux/ HNS pSB1C3 + lux experiment
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | Accomplishments
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Website
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Make LB Media
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | chlor
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | amp
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | chlor + amp
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | kan
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | To Make Glycerol stocks:
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up SY102 in Chlorom to isolate CDE
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up SY104 in Chlorom to isolate CDE
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up SY102 in Amp to isolate AB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up SY102 in Amp + Chlorom to isolate split lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up SY104 in Amp + Chlorom to isolate split lux
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | To see if plasmid has been lost
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up SY102 plated on Chlor in Amp to see if it survives
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up SY104 plated on Chlor in Amp to see if it survives
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Grow up SY102 plated on Amp in Chlor to see if it survives
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | </ul>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | For the timecourse experiment:
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Take the following upstairs:
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pSB1C3 + lux in HNS (x2)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pSB1C3 + lux in NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in HNS (x2)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn + mRFP in BL21 → should be in foil
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn + mRFP in DH5alpha→ should be in foil
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Take aluminum foil upstairs just in case you cannot find it.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At 9PM,
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Measure OD of cultures below to ensure that they have reached saturation (see if one of the two tubes has not grown properly)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in HNS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in HNS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 in 100 ml media culture flasks (so we add 1000 ul of culture into the appropriate flask)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pSB1C3 + lux in HNS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pSB1C3 + lux in NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in HNS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 in 20 ml media culture flasks(so we add 200 ul of culture into the appropriate flask) COVER GIANT FLASKS IN FOIL
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn + mRFP in BL21 → should be in foil
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pDawn + mRFP in DH5alpha→ should be in foil
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Place all flasks in the incubator; make sure they are closed; make sure that the flasks are not falling out
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At 11:30-12:00PM,
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Measure the OD of ALL cultures; make sure you use conversion to make sure it is appropriate to induce.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Separate the following cultures into three tubes with 5 ml of liquid in each one. Then add 50 ul of arabinose to each tube.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pSB1C3 + lux in HNS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pSB1C3 + lux in NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | pACBB + lux in HNS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Also add arabinose to the big flasks and place them in teh incubator
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Clean the inside of the culture tube with Karol’s set up with isoproponol. Add 3 mL of the appropriate culture into each tube. Turn on the
| + | |
| − | light by making sure the switch is in - position and that the yellow is connected to red and black is connected to black.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Once the light turns on, cover the lids with aluminum foil and make sure controls are also properly covered.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ul>
| + | |
| − | </ul>
| + | |
| − | <p dir="ltr">
| + | |
| − | Aims for Tomorrow:
| + | |
| − | </p>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Sender-cell trendlines:
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Start 10-17 hour luminescence measurement for pSB1C3 + lux in both HNS and NEB10
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Analyze the data with the pDawn + mRFP experiment. Determine the threshold?
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 (at 10:30) pSB1C3 + lux in HNS (in LB + chlorom) to observe the trend in hours 1-6
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At 12:30 PM
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Separate into 4 different tubes with 3 mL each (1 is an uninduced control)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Add 30 ul of 1 M arabinose to each tube and take OD and luminescence measurements (approximately 1 PM - 7 PM)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Housekeeping:
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Glycerol stock SY102 and SY104 + add to LIMS
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pick up plates and nalidixic acid from the cell center.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Pour LB + Chlor plates
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Plan co-tubing experiment:
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | 1:100 pSB1C3 + lux in HNS_1000 uL to 100 mL; 1:100 pDawn + mRFP in DH5alpha (in the dark)__500 uL to 50 mL
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Prepare 6 tubes
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Tube 1 (X2)- in dark; IND 10 mL of pSB1C3 + lux; 3 mL of pDawn + mRFP inside
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Tube 2 (X2)- in dark; UNIND 10 mL of pSB1C3 + lux; 3 mL of pDawn + mRFP inside
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Tube 3 (X2)- in light; 3 mL of pDawn + mRFP inside
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At 2 hours, check the OD has reached ~0.3-0.4. Add the appropriate volume of the appropriate culture to the tubes after inducing with 1 M
| + | |
| − | arabinose.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | At 20 hr take a fluorescence measurement.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Replating for split lux (yesterday proved that the cells have not lost the plasmid)
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <ol>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Re-plate SY102 in Chlorom to isolate CDE
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Re-plate SY104 in Chlorom to isolate CDE
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Re-plate SY102 in Amp to isolate AB
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | <li dir="ltr">
| + | |
| − | <p dir="ltr">
| + | |
| − | Tomorrow grow up all plates in opposite antibiotic.
| + | |
| − | </p>
| + | |
| − | </li>
| + | |
| − | </ol>
| + | |
| − | </ol>
| + | |
| − | <br/>
| + | |
| − | <p dir="ltr">
| + | |
| − | 8/19/15
| + | |
| − | </p>
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| − | Aims for Today:
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| − | Sender-cell trendlines:
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| − | Start 10-17 hour luminescence measurement for pSB1C3 + lux in both HNS and NEB10
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| − | Analyze the data with the pDawn + mRFP experiment. Determine the threshold?
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| − | </ol>
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| − | Housekeeping:
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| − | </p>
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| − | <ol>
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| − | Glycerol stock SY102 and SY104 + add to LIMS
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| − | Pick up plates and nalidixic acid from the cell center.
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| − | Pour LB + Chlor plates
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| − | Plan co-tubing experiment:
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| − | <ol>
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| − | 1:100 pSB1C3 + lux in HNS_1000 uL to 100 mL; 1:100 pDawn + mRFP in DH5alpha (in the dark)__1000 uL to 100 mL
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| − | Prepare 6 tubes
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| − | Tube 1 (X2)- in dark; IND 10 mL of pSB1C3 + lux; 3 mL of pDawn + mRFP inside
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| − | Tube 2 (X2)- in dark; UNIND 10 mL of pSB1C3 + lux; 3 mL of pDawn + mRFP inside
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| − | Tube 3 (X2)- in light; 3 mL of pDawn + mRFP inside
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| − | </p>
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| − | </ol>
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| − | At 2 hours, check the OD has reached ~0.3-0.4. Add the appropriate volume of the appropriate culture to the tubes after inducing with 1 M
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| − | arabinose.
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| − | At 20 hr take a fluorescence measurement.
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| − | </p>
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| − | </ol>
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| − | Replating for split lux (yesterday proved that the cells have not lost the plasmid)
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| − | </p>
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| − | <ol>
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| − | Re-plate SY102 in Chlorom to isolate CDE
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| − | </p>
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| − | </li>
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| − | Re-plate SY104 in Chlorom to isolate CDE
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| − | </p>
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| − | Re-plate SY102 in Amp to isolate AB
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| − | </p>
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| − | Tomorrow grow up all plates in opposite antibiotic.
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| − | </p>
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| − | </ol>
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| − | <p dir="ltr">
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| − | For Jane,
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| − | </p>
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| − | <ol>
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| − | <p dir="ltr">
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| − | Grow up
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| − | </p>
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| − | <ol>
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| − | <p dir="ltr">
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| − | HNS
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| − | </p>
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| − | <p dir="ltr">
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| − | Split lux (both strains)
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| − | </p>
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| − | </li>
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| − | <p dir="ltr">
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| − | Make sure she has enough flasks
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| − | </p>
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| − | 1:100 at 6 AM to 7 AM ??? so induce at 8 AM - 9AM
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| − | </p>
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| − | </li>
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| − | <li dir="ltr">
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| − | <p dir="ltr">
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| − | Make sure she has 1 M arabinose and appropriate nalidixic acid solution made up
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| − | </p>
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| − | </li>
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| − | <p dir="ltr">
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| − | Split up the timepoints so that she doesn’t have to do everything by herself
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| − | </p>
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| − | </li>
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| − | </ol>
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| − | </ol>
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| − | <p dir="ltr">
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| − | Website and poster layout completed.
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| − | </p>
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| − | </li>
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| − | </ol>
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| − | <br/>
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| | </body> | | </body> |
| | </html> | | </html> |
