Difference between revisions of "Team:UCLA/Notebook/Honeybee Silk/11 May 2015"

 
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  <summary>May 11 Sequencing of silk biobricks</summary>
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        <p>Today we sent in both our constructs to GeneWiz for sequencing. </p>
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            <li> For sequencing primers, I used the igem vf2 primers and vr primers that bind to the psb1c3 backbone, as well as a sequencing primer that we designed to bind to the silk sequence (p13) see https://benchling.com/s/p7bClpzQ/edit </li>
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            <li>The sequencing results came back and checked out for sample 1 of construct 1 (silk coding region) and samples 1 and 2 for construc 2 (regulatory elements + silk) <b> i.e. we have our first honeybee biobricks!!! </b> </li>
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            <li> For sequencing results of construct 1 see https://benchling.com/s/4XT6VRRU/edit (click on alignments on the right side of screen) </li>
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            <li> For sequencing results of construct 2 see https://benchling.com/s/TxD8z7Kq/edit (click on alignments on the right side of screen) </li>
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<ul>
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<a href="https://2015.igem.org/Team:UCLA"><li>HOME</li></a>
 +
 
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<a href="https://2015.igem.org/Team:UCLA/Team"><li>TEAM</li></a>
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<a href="#"><li>PROJECTS
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            <ul>
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<!-- <a href="https://2015.igem.org/Team:UCLA/Description"><li>Description</li></a>
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<a href="https://2015.igem.org/Team:UCLA/Experiments"><li>Experiments &amp; Protocols</li></a>  
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<a href="https://2015.igem.org/Team:UCLA/Results"><li>Results</li></a> 
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<a href="https://2015.igem.org/Team:UCLA/Design"><li>Design</li></a>-->
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<a href="https://2015.igem.org/Team:UCLA/Projects/Silks"><li>Silk Genetics</li></a>
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<a href="https://2015.igem.org/Team:UCLA/Projects/Functionalization"><li>Functionalization</li></a>
 +
<a href="https://2015.igem.org/Team:UCLA/Projects/Materials"><li>Materials Processing</li></a>
 +
<a href="https://2015.igem.org/Team:UCLA/Projects/Protein_Cages"><li>Protein Cages</li></a>
 +
</ul>
 +
</li></a>
 +
 
 +
<a href="#"><li>PARTS
 +
            <ul>
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<a href="https://2015.igem.org/Team:UCLA/Parts"><li>Team Parts</li></a>
 +
<a href="https://2015.igem.org/Team:UCLA/Basic_Part"><li>Basic Parts</li></a>  
 +
<a href="https://2015.igem.org/Team:UCLA/Composite_Part"><li>Composite Parts</li></a>
 +
<a href="https://2015.igem.org/Team:UCLA/Part_Collection"><li>Part Collection</li></a> 
 +
</ul>
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</li></a>
 +
 
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<a href="https://2015.igem.org/Team:UCLA/Notebook"><li>NOTEBOOKS
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                                                <ul>
 +
<a href="https://2015.igem.org/Team:UCLA/Notebook/Spider_Silk_Genetics"><li>Spider Silk Genetics</li></a>
 +
<a href="https://2015.igem.org/Team:UCLA/Notebook/Honeybee_Silk"><li>Honeybee Silk</li></a>  
 +
<a href="https://2015.igem.org/Team:UCLA/Notebook/Recombinant_Expression"><li>Recombinant Silk Functionalization</li></a> 
 +
<a href="https://2015.igem.org/Team:UCLA/Notebook/Materials"><li>Materials  Processing</li></a> 
 +
<a href="https://2015.igem.org/Team:UCLA/Notebook/Protein_Cages"><li>Protein Cages</li></a>
 +
                                                        <a href="https://2015.igem.org/Team:UCLA/Notebook/Interlab_Study"><li>UCLA Measurement Interlab Study</li></a>
 +
</ul>
 +
                                        </li></a>
 +
 
 +
<a href="#"><li>MEETING NOTES
 +
                                                <ul>
 +
<a href="https://2015.igem.org/Team:UCLA/Notebook/General_Meetings"><li>General Meetings</li></a>
 +
                                                        <a href="https://2015.igem.org/Team:UCLA/Notebook/Coordinator_Meetings"><li>Coordinator Meetings</li></a>
 +
<a href="https://2015.igem.org/Team:UCLA/Notebook/Advisor_Meetings"><li>Advisor Meetings</li></a>   
 +
</ul>
 +
                                        </li></a>
 +
 
 +
     
 +
<a href="https://2015.igem.org/Team:UCLA/Attributions"><li>ATTRIBUTIONS</li></a>
 +
 
 +
<a href="https://2015.igem.org/Team:UCLA/Collaborations"><li>COLLABORATIONS</li></a>
 +
 
 +
<a href="https://2015.igem.org/Team:UCLA/Practices"><li>HUMAN PRACTICES</li></a>
 +
 
 +
<a href="https://2015.igem.org/Team:UCLA/Safety"><li>SAFETY</li></a>
 +
 
 +
<a href="https://2015.igem.org/Team:UCLA/Modeling"><li>MODELING</li></a>
 +
 
 +
<a href="https://2015.igem.org/Team:UCLA/Measurement"><li>MEASUREMENT STUDY</li></a>
 +
 
 +
<a href="https://2015.igem.org/Team:UCLA/Software"><li>SOFTWARE</li></a>
 +
 
 +
<a href="https://2015.igem.org/Team:UCLA/Entrepreneurship"><li>ENTREPRENEURSHIP</li></a>
 +
 
 +
</ul>
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</div>
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='''Inoculation of 5/10 Transformation'''=
 +
 +
*Transformation was good
 +
*I picked three colonies from the 1:1 kanamycin plate and suspended each in 100uL ddH2O (labeled pET 1, 2, and 3 respectively)
 +
*Each sample was inoculated in 3 different culture tubes with 5mL LB broth and 5uL 1000X kanamycin and was incubated at 37C.
 +
 +
='''Sequencing of Silk Biobricks'''=
 +
 +
We sent in both of our constructs to Genewiz for sequencing.
 +
 +
*For sequencing primers, we used the iGEM VF2 and VR primers that bind to the psb1c3 backbone, as well as the sequencing primer that we designed to bind to the silk sequence (p13) see https://benchling.com/s/p7bClpzQ/edit
 +
*The sequencing results can back and checked out for sample 1 of construct 1 (silk coding region) and samples 1 and 2 for construct 2 (regulartory elements+silk)
 +
**We have our first honeybee biobricks!
 +
*For sequencing results of construct 1 see  https://benchling.com/s/4XT6VRRU/edit (click on alignments on the right side of screen)
 +
*For sequencing results of construct 2 see https://benchling.com/s/TxD8z7Kq/edit (click on alignments on the right side of screen)

Latest revision as of 21:05, 10 July 2015

Inoculation of 5/10 Transformation

  • Transformation was good
  • I picked three colonies from the 1:1 kanamycin plate and suspended each in 100uL ddH2O (labeled pET 1, 2, and 3 respectively)
  • Each sample was inoculated in 3 different culture tubes with 5mL LB broth and 5uL 1000X kanamycin and was incubated at 37C.

Sequencing of Silk Biobricks

We sent in both of our constructs to Genewiz for sequencing.

  • For sequencing primers, we used the iGEM VF2 and VR primers that bind to the psb1c3 backbone, as well as the sequencing primer that we designed to bind to the silk sequence (p13) see https://benchling.com/s/p7bClpzQ/edit
  • The sequencing results can back and checked out for sample 1 of construct 1 (silk coding region) and samples 1 and 2 for construct 2 (regulartory elements+silk)
    • We have our first honeybee biobricks!
  • For sequencing results of construct 1 see https://benchling.com/s/4XT6VRRU/edit (click on alignments on the right side of screen)
  • For sequencing results of construct 2 see https://benchling.com/s/TxD8z7Kq/edit (click on alignments on the right side of screen)