Difference between revisions of "Team:Penn/Notebook"

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       6/9/2015
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       6/8/2015
 
       <div class="arrow-down"></div>
 
       <div class="arrow-down"></div>
 
   </h2>
 
   </h2>
 
   <div class="wrapper">
 
   <div class="wrapper">
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<p><strong>Aims for today:</strong></p>
+
    <div class="content">
 +
    <p><strong>Aims for the day:</strong></p>
 
<ol>
 
<ol>
−
<li>PCR cleanup for lacZ</li>
+
<li>Run a gel after digestion of lacZ and pDawn</li>
−
<li>Digest lacZ and pDawn</li>
+
<li>Gel extraction &amp; digest cleanup</li>
−
<li>Digest cleanup</li>
+
<li>Ligation</li>
−
<li>Ligation of pDawn and lacZ</li>
+
<li>Transformation</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>
+
 
</ol>
 
</ol>
 
<p>&nbsp;</p>
 
<p>&nbsp;</p>
 
<p><strong>Accomplishments:</strong></p>
 
<p><strong>Accomplishments:</strong></p>
−
<p>&nbsp;</p>
 
 
<ol>
 
<ol>
−
<li>PCR cleanup for lacZ</li>
+
<li>Ran a gel after digestion</li>
−
<li>Digest of lacZ and pDawn</li>
+
<li>Performed gel extraction and digestion cleanup</li>
−
<li>Digest cleanup of lacZ and pDawn</li>
+
<li>Measured concentrations of pDawn and lacZ with nanodrop</li>
−
<li>Ligation</li>
+
−
<li>Lux box transformation</li>
+
−
<li>Began working on plasmid design</li>
+
−
<li>Inoculated pDawn</li>
+
 
</ol>
 
</ol>
 
<p>&nbsp;</p>
 
<p>&nbsp;</p>
−
<p><strong>&nbsp;Aims for tomorrow:</strong></p>
+
<p><strong>Aims for tomorrow:</strong></p>
 
<ol>
 
<ol>
−
<li>Potentially design primers for cloning Lux box into pDawn/lacZ</li>
+
<li>PCR clean-up for new PCR of lacZ</li>
−
<li>Miniprep pDawn</li>
+
<li>Digestion of lacZ and pDawn with new Nde1 and BamHI</li>
−
<li>Design SapI primers</li>
+
<li>Digest clean-up</li>
−
<li>Design compatibility system</li>
+
<li>Ligation of lacZ and pDawn</li>
−
<li>Pick colonies for BioBrick, inoculate, glycerol stock</li>
+
<li>Lux box transformation into E. coli</li>
−
<li>Colony PCR pDawn + lacZ, glycerol stock, miniprep, sequencing</li>
+
−
<li>Induce lux box w/ arabinose</li>
+
 
</ol>
 
</ol>
−
<p>&nbsp;</p>
+
    </div>
−
<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>
+
−
<p>&nbsp;</p>
+
−
    <div class="content">
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   </div>
 
   </div>
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Revision as of 01:50, 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

6/8/2015

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