Difference between revisions of "Team:Penn/Notebook"

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       6/9/2015
 
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<p><strong>Aims for today:</strong></p>
 +
<ol>
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<li>PCR cleanup for lacZ</li>
 +
<li>Digest lacZ and pDawn</li>
 +
<li>Digest cleanup</li>
 +
<li>Ligation of pDawn and lacZ</li>
 +
<li>Lux box transformation</li>
 +
<li>Design one plasmid system with Lux box/lacZ and design two plasmid system, one with Lux box and the other with the lacZ reporter</li>
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</ol>
 +
<p>&nbsp;</p>
 +
<p><strong>Accomplishments:</strong></p>
 +
<p>&nbsp;</p>
 +
<ol>
 +
<li>PCR cleanup for lacZ</li>
 +
<li>Digest of lacZ and pDawn</li>
 +
<li>Digest cleanup of lacZ and pDawn</li>
 +
<li>Ligation</li>
 +
<li>Lux box transformation</li>
 +
<li>Began working on plasmid design</li>
 +
<li>Inoculated pDawn</li>
 +
</ol>
 +
<p>&nbsp;</p>
 +
<p><strong>&nbsp;Aims for tomorrow:</strong></p>
 +
<ol>
 +
<li>Potentially design primers for cloning Lux box into pDawn/lacZ</li>
 +
<li>Miniprep pDawn</li>
 +
<li>Design SapI primers</li>
 +
<li>Design compatibility system</li>
 +
<li>Pick colonies for BioBrick, inoculate, glycerol stock</li>
 +
<li>Colony PCR pDawn + lacZ, glycerol stock, miniprep, sequencing</li>
 +
<li>Induce lux box w/ arabinose</li>
 +
</ol>
 +
<p>&nbsp;</p>
 +
<p><strong>Questions:</strong></p>
 +
<p>In the &ldquo;empty&rdquo; region of pDawn (the only space we think a biobrick can be cloned) there&rsquo;s only one restriction enzyme that cuts once in that region and nowhere else, SapI.</p>
 +
<ol>
 +
<li>Are we right in assuming that empty space is the only place we can clone in a biobrick?</li>
 +
<li>Would it be okay if we used only one restriction enzyme for the ligation instead of two?</li>
 +
</ol>
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<p>&nbsp;</p>
 
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Revision as of 01:48, 17 September 2015

University of Pennsylvania iGEM

Content Expander

expanding content since 2014

robsawyer.me

6/8/2015

Aims for the day:

  1. Run a gel after digestion of lacZ and pDawn
  2. Gel extraction & digest cleanup
  3. Ligation
  4. Transformation

 

Accomplishments:

  1. Ran a gel after digestion
  2. Performed gel extraction and digestion cleanup
  3. Measured concentrations of pDawn and lacZ with nanodrop

 

Aims for tomorrow:

  1. PCR clean-up for new PCR of lacZ
  2. Digestion of lacZ and pDawn with new Nde1 and BamHI
  3. Digest clean-up
  4. Ligation of lacZ and pDawn
  5. Lux box transformation into E. coli

6/9/2015

Aims for today:

  1. PCR cleanup for lacZ
  2. Digest lacZ and pDawn
  3. Digest cleanup
  4. Ligation of pDawn and lacZ
  5. Lux box transformation
  6. Design one plasmid system with Lux box/lacZ and design two plasmid system, one with Lux box and the other with the lacZ reporter

 

Accomplishments:

 

  1. PCR cleanup for lacZ
  2. Digest of lacZ and pDawn
  3. Digest cleanup of lacZ and pDawn
  4. Ligation
  5. Lux box transformation
  6. Began working on plasmid design
  7. Inoculated pDawn

 

 Aims for tomorrow:

  1. Potentially design primers for cloning Lux box into pDawn/lacZ
  2. Miniprep pDawn
  3. Design SapI primers
  4. Design compatibility system
  5. Pick colonies for BioBrick, inoculate, glycerol stock
  6. Colony PCR pDawn + lacZ, glycerol stock, miniprep, sequencing
  7. Induce lux box w/ arabinose

 

Questions:

In the “empty” region of pDawn (the only space we think a biobrick can be cloned) there’s only one restriction enzyme that cuts once in that region and nowhere else, SapI.

  1. Are we right in assuming that empty space is the only place we can clone in a biobrick?
  2. Would it be okay if we used only one restriction enzyme for the ligation instead of two?