Difference between revisions of "Team:Georgia State/notebookdata.html"

 
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<div id="date-6-10">
 
<div id="date-6-10">
 
   <h1>Tuesday, June 10th.</h1>
 
   <h1>Tuesday, June 10th.</h1>
−
   <ul>
+
   </div>
−
    <li>Revision of the protocols for next month&rsquo;s lab-work, 12:00.</li>
+
−
    <ul>
+
−
      <li>Everyone.</li>
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−
    </ul>
+
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    <li>Revision of human practice ideas and collaborations with other teams, 13:00.</li>
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−
    <ul>
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−
      <li>Everyone.</li>
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−
    </ul>
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−
  </ul>
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−
</div>
+
  
 
<div id="date-6-11">
 
<div id="date-6-11">
 
   <h1>Wednesday, June 11th.</h1>
 
   <h1>Wednesday, June 11th.</h1>
−
   <ul>
+
   </div>
−
    <li>Electro-competent cells, 13:00.</li>
+
−
    <ul>
+
−
      <li>Cinthya</li>
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−
    </ul>
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−
  </ul>
+
−
</div>
+
  
 
<div id="date-6-12">
 
<div id="date-6-12">
 
   <h1>Thursday, June 12th.</h1>
 
   <h1>Thursday, June 12th.</h1>
−
   <ul>
+
   </div>
−
    <li>Sponsorships, all day.</li>
+
−
    <ul>
+
−
      <li>Cinthya.</li>
+
−
    </ul>
+
−
  </ul>
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−
</div>
+
  
 
<div id="date-6-13">
 
<div id="date-6-13">
 
   <h1>Friday, June 13th.</h1>
 
   <h1>Friday, June 13th.</h1>
−
   <ul>
+
   </div>
−
    <li>Meeting with advisors, 17:00.</li>
+
−
    <li>Calcium-competent cells, 13:00.</li>
+
−
    <ul>
+
−
      <li>Cinthya.</li>
+
−
    </ul>
+
−
  </ul>
+
−
</div>
+
  
 
<div id="date-6-14">
 
<div id="date-6-14">
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<div id="date-6-16">
 
<div id="date-6-16">
 
   <h1>Monday, June 16th.</h1>
 
   <h1>Monday, June 16th.</h1>
−
   <ul>
+
   </div>
−
    <li>Electro-competent cells, 14:00.</li>
+
−
    <ul>
+
−
      <li>Cinthya, Mercedes.</li>
+
−
    </ul>
+
−
    <li>Minipreps, 16:00.</li>
+
−
    <ul>
+
−
      <li>Cinthya.</li>
+
−
    </ul>
+
−
    <li>Electrophoresis, 19:00.</li>
+
−
    <ul>
+
−
      <li>Cinthya.</li>
+
−
    </ul>
+
−
    <li>Inoculations, 20:00.</li>
+
−
    <ul>
+
−
      <li>Mercedes.</li>
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−
    </ul>
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−
  </ul>
+
−
</div>
+
  
 
<div id="date-6-17">
 
<div id="date-6-17">
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<div id="date-6-18">
 
<div id="date-6-18">
 
   <h1>Wednesday, June 18th.</h1>
 
   <h1>Wednesday, June 18th.</h1>
−
   <ul>
+
   </div>
−
    <li>Electro-competent cells, 10:00.</li>
+
−
    <ul>
+
−
      <li>Cinthya.</li>
+
−
    </ul>
+
−
    <li>Minipreps, 13:00.</li>
+
−
    <ul>
+
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
    <li>Electrophoresis, 16:00.</li>
+
−
    <ul>
+
−
      <li>Cinthya.</li>
+
−
    </ul>
+
−
    <li>Inoculations, 20:00.</li>
+
−
    <ul>
+
−
      <li>Cinthya.</li>
+
−
    </ul>
+
−
  </ul>
+
−
</div>
+
  
 
<div id="date-6-19">
 
<div id="date-6-19">
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<div id="date-6-20">
 
<div id="date-6-20">
 
   <h1>Friday, June 20th.</h1>
 
   <h1>Friday, June 20th.</h1>
−
   <ul>
+
   </div>
−
    <li>Meeting with advisors, 17:00.</li>
+
−
  </ul>
+
−
</div>
+
  
 
<div id="date-6-21">
 
<div id="date-6-21">
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<div id="date-6-23">
 
<div id="date-6-23">
 
   <h1>Monday, June 23rd.</h1>
 
   <h1>Monday, June 23rd.</h1>
−
   <ul>
+
    
−
    <li>Calcium-competent cells, 12:00.</li>
+
−
    <ul>
+
−
      <li>Cinthya.</li>
+
−
    </ul>
+
−
  </ul>
+
 
</div>
 
</div>
  
 
<div id="date-6-24">
 
<div id="date-6-24">
 
   <h1>Tuesday, June 24th.</h1>
 
   <h1>Tuesday, June 24th.</h1>
−
   <ul>
+
    
−
    <li>Minipreps, 11:00</li>
+
−
    <ul>
+
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
    <li>Electrophoresis, 14:00.</li>
+
−
    <ul>
+
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
    <li>Inoculations. 18:00.</li>
+
−
    <ul>
+
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
  </ul>
+
 
</div>
 
</div>
  
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<div id="date-6-26">
 
<div id="date-6-26">
 
   <h1>Thursday, June 26th.</h1>
 
   <h1>Thursday, June 26th.</h1>
−
  <ul>
+
 
−
    <li>Minipreps, 10:00</li>
+
 
−
    <ul>
+
    
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
    <li>Electrophoresis, 13:30.</li>
+
−
    <ul>
+
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
   </ul>
+
 
</div>
 
</div>
  
 
<div id="date-6-27">
 
<div id="date-6-27">
 
   <h1>Friday, June 27th.</h1>
 
   <h1>Friday, June 27th.</h1>
−
  <ul>
+
<li> Cara Jones</li>
−
    <li>Meeting with advisors, 17:00.</li>
+
<ul>Plasmid Resuspension, Primer Dilutions, PCR: <a href="https://static.igem.org/mediawiki/2017/b/b7/T--Georgia_State--6_27_17_cj.pdf"> See attached procedure for buffer recipes and instructions.</a>  
−
  </ul>
+
  </div>
−
</div>
+
  
 
<div id="date-6-28">
 
<div id="date-6-28">
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   <h1>Friday, July 4th.</h1>
 
   <h1>Friday, July 4th.</h1>
 
   <ul>
 
   <ul>
−
    <li>Minipreps, 12:00.</li>
+
    
−
    <ul>
+
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
    <li>Practice: Ligations, 15:00.</li>
+
−
    <ul>
+
−
      <li>Eduardo Ramirez (1).</li>
+
−
    </ul>
+
−
    <li>Practice: Electrophoresis, 17:00.</li>
+
−
    <ul>
+
−
      <li>Eduardo Ramirez (1)</li>
+
−
    </ul>
+
−
   </ul>
+
 
</div>
 
</div>
  
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<div id="date-7-6">
 
<div id="date-7-6">
 
   <h1>Thursday, July 6th.</h1>
 
   <h1>Thursday, July 6th.</h1>
 +
<li> Cara Jones</li>
 +
<ul>Ligation of Mamba insert+pGEX vector and DNA clean up of pGEX+mamba restriction digest: <a href="https://static.igem.org/mediawiki/2017/0/0d/T--Georgia_State--7_6_17_cj.pdf"> See attached procedure for instructions.</a>
 
</div>
 
</div>
  
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<ul><b>Procedure:</b><a href="https://static.igem.org/mediawiki/2017/0/0a/T--Georgia_State--APTestprotocol.pdf">See attached procedure.</a></ul>   
 
<ul><b>Procedure:</b><a href="https://static.igem.org/mediawiki/2017/0/0a/T--Georgia_State--APTestprotocol.pdf">See attached procedure.</a></ul>   
 
<ul><b>Results:</b> The Ap spread outward but stayed confined meaning the crosslinking worked. Adding the BCIP solution to the tube with the paper produced a noticeable discoloration. Adding the enzyme to the BCIP only produced a small discoloration. The binding of the alkaline phosphatase and BCIP to the paper was successful. The reaction worked only in the intended areas meaning that if the AP is bound to the factor c it may produce a signal only if it is cut, the AP nor BCIP disperses in the solution after being bound to the paper.<br><img src="https://static.igem.org/mediawiki/2017/c/c5/T--Georgia_State--APTest.png"></ul>
 
<ul><b>Results:</b> The Ap spread outward but stayed confined meaning the crosslinking worked. Adding the BCIP solution to the tube with the paper produced a noticeable discoloration. Adding the enzyme to the BCIP only produced a small discoloration. The binding of the alkaline phosphatase and BCIP to the paper was successful. The reaction worked only in the intended areas meaning that if the AP is bound to the factor c it may produce a signal only if it is cut, the AP nor BCIP disperses in the solution after being bound to the paper.<br><img src="https://static.igem.org/mediawiki/2017/c/c5/T--Georgia_State--APTest.png"></ul>
 +
<li> Cara Jones</li>
 +
<ul>Colony PCR: <a href="https://static.igem.org/mediawiki/2017/2/20/T--Georgia_State--7_7_17_cj.pdf"> See attached procedure for instructions.</a>
 
      
 
      
 
</div>
 
</div>
  
 
<div id="date-7-8">
 
<div id="date-7-8">
−
   <h1>Tuesday, July 8th.</h1>
+
   <h1>Saturday, July 8th.</h1>
 
</div>
 
</div>
  
 
<div id="date-7-9">
 
<div id="date-7-9">
−
   <h1>Wednesday, July 9th.</h1>
+
   <h1>Sunday, July 9th.</h1>
 
</div>
 
</div>
  
 
<div id="date-7-10">
 
<div id="date-7-10">
−
   <h1>Thursday, July 10th.</h1>
+
   <h1>Monday, July 10th.</h1>
−
  <ul>
+
<li> Cara Jones</li>
−
    <li>Practice: Transformation of calcium-competent cells, 11:00,</li>
+
<ul>Restriction digest of mamba in pGEX: <a href="https://static.igem.org/mediawiki/2017/e/ed/T--Georgia_State--7_10_17_cj.pdf"> See attached procedure for instructions.</a>  
−
    <ul>
+
 
−
      <li>Eduardo Ramirez (1), Claudia.</li>
+
 
−
    </ul>
+
−
    <li>Inoculation on LB-agar plates, 13:00.</li>
+
−
    <ul>
+
−
      <li>Claudia.</li>
+
−
    </ul>
+
−
    <li>Laboratory summer cleaning, 15:00.</li>
+
−
    <ul>
+
−
      <li>Claudia.</li>
+
−
    </ul>
+
−
  </ul>
+
 
</div>
 
</div>
  
 
<div id="date-7-11">
 
<div id="date-7-11">
−
   <h1>Friday, July 11th.</h1>
+
   <h1>Tuesday, July 11th.</h1>
 +
<li> Cara Jones</li>
 +
<ul>Sequencing results from macrogen and planning : <a href="https://static.igem.org/mediawiki/2017/c/c7/T--Georgia_State--7_11_17_cj.pdf"> See attached.</a>  
 
</div>
 
</div>
  
 
<div id="date-7-12">
 
<div id="date-7-12">
−
   <h1>Saturday, July 12th.</h1>
+
   <h1>Wednesday, July 12th.</h1>
 
</div>
 
</div>
  
 
<div id="date-7-13">
 
<div id="date-7-13">
−
   <h1>Sunday, July 13th.</h1>
+
   <h1>Thursday, July 13th.</h1>
 +
<li> Cara Jones</li>
 +
<ul>Transformation of mamba in pGEX into Bl21 Competent cells: <a href="https://static.igem.org/mediawiki/2017/5/5a/T--Georgia_State--7_13_17_cj.pdf"> See attached procedure for instructions.</a>  
 
</div>
 
</div>
  
 
<div id="date-7-14">
 
<div id="date-7-14">
−
   <h1>Monday, July 14th.</h1>
+
   <h1>Friday, July 14th.</h1>
 
</div>
 
</div>
  
 
<div id="date-7-15">
 
<div id="date-7-15">
−
   <h1>Tuesday, July 15th.</h1>
+
   <h1>Saturday, July 15th.</h1>
−
   <ul>
+
   </div>
−
    <li>Practice: taking DNA out of the iGEM plates, 13:00.</li>
+
−
    <ul>
+
−
      <li>Claudia.</li>
+
−
    </ul>
+
−
    <li>Transformation with calcium-competent cells, 13:30.</li>
+
−
    <ul>
+
−
      <li>Claudia.</li>
+
−
    </ul>
+
−
    <li>Inoculations, 15:00.</li>
+
−
    <ul>
+
−
      <li>Claudia.</li>
+
−
    </ul>
+
−
  </ul>
+
−
</div>
+
  
 
<div id="date-7-16">
 
<div id="date-7-16">
−
   <h1>Wednesday, July 16th.</h1>
+
   <h1>Sunday, July 16th.</h1>
−
   <ul>
+
   </div>
−
    <li>Minipreps, 12:00.</li>
+
−
    <ul>
+
−
      <li>Eduardo Ramirez (1).</li>
+
−
    </ul>
+
−
    <li>Ligations, 15:00.</li>
+
−
    <ul>
+
−
      <li>Eduardo Ramirez (1).</li>
+
−
    </ul>
+
−
    <li>Electrophoresis, 18:00.</li>
+
−
    <li>Practice: DNA recovery from gel, 20:00.</li>
+
−
    <ul>
+
−
      <li>Eduardo Ramirez (1)</li>
+
−
    </ul>
+
−
  </ul>
+
−
</div>
+
  
 
<div id="date-7-17">
 
<div id="date-7-17">
−
   <h1>Thursday, July 17th.</h1>
+
   <h1>Monday, July 17th.</h1>
 +
<li> Cara Jones</li>
 +
<ul>Interlab 2017 transformations and Mamba in pGEX protein expression: <a href="https://static.igem.org/mediawiki/2017/c/c7/T--Georgia_State--7_17_17_cj.pdf"> See attached procedure for instructions.</a>  
 
</div>
 
</div>
  
 
<div id="date-7-18">
 
<div id="date-7-18">
−
   <h1>Friday, July 18th.</h1>
+
   <h1>Tuesday, July 18th.</h1>
 
</div>
 
</div>
  
 
<div id="date-7-19">
 
<div id="date-7-19">
−
   <h1>Saturday, July 19th.</h1>
+
   <h1>Wednesday, July 19th.</h1>
−
   <ul>
+
   </div>
−
    <li>Meeting with advisors to talk about gene problems and doubts, 17:00.</li>
+
−
  </ul>
+
−
</div>
+
  
 
<div id="date-7-20">
 
<div id="date-7-20">
−
   <h1>Sunday, July 20th.</h1>
+
   <h1>Thursday, July 20th.</h1>
 
</div>
 
</div>
  
 
<div id="date-7-21">
 
<div id="date-7-21">
−
   <h1>Monday, July 21st.</h1>
+
   <h1>Friday, July 21st.</h1>
 
</div>
 
</div>
  
 
<div id="date-7-22">
 
<div id="date-7-22">
−
   <h1>Tuesday, July 22nd.</h1>
+
   <h1>Saturday, July 22nd.</h1>
−
   <ul>
+
   </div>
−
    <li>Minipreps, 11:00.</li>
+
−
    <ul>
+
−
      <li>Claudia.</li>
+
−
    </ul>
+
−
    <li>Ligations, 15:00.</li>
+
−
    <ul>
+
−
      <li>Claudia.</li>
+
−
    </ul>
+
−
    <li>Electrophoresis, 19:00.</li>
+
−
    <ul>
+
−
      <li>Claudia</li>
+
−
    </ul>
+
−
    <li>Practice: DNA recovery from gel, 21:00.</li>
+
−
    <ul>
+
−
      <li>Claudia.</li>
+
−
    </ul>
+
−
  </ul>
+
−
</div>
+
  
 
<div id="date-7-23">
 
<div id="date-7-23">
−
   <h1>Wednesday, July 23rd.</h1>
+
   <h1>Sunday, July 23rd.</h1>
 
</div>
 
</div>
  
 
<div id="date-7-24">
 
<div id="date-7-24">
−
   <h1>Thursday, July 24th.</h1>
+
   <h1>Monday, July 24th.</h1>
 
</div>
 
</div>
  
 
<div id="date-7-25">
 
<div id="date-7-25">
−
   <h1>Friday, July 25th.</h1>
+
   <h1>Tuesday, July 25th.</h1>
 
</div>
 
</div>
  
 
<div id="date-7-26">
 
<div id="date-7-26">
−
   <h1>Saturday, July 26th.</h1>
+
   <h1>Wednesday, July 26th.</h1>
−
   <ul>
+
   </div>
−
    <li>Meeting with advisors, 17:00.</li>
+
    
−
   </ul>
+
−
  <h1><strong><strong>&nbsp;</strong></strong></h1>
+
−
  <h1>Monday, July 29th.</h1>
+
−
  <ul>
+
−
    <li>Meeting to plan the semester&rsquo;s activities, 14:00.</li>
+
−
    <ul>
+
−
      <li>Schedule of people in the lab.</li>
+
−
      <li>Ways to integrate other students.</li>
+
−
      <li>Events to talk about iGEM.</li>
+
−
    </ul>
+
−
  </ul>
+
−
</div>
+
−
 
+
 
<div id="date-7-27">
 
<div id="date-7-27">
−
   <h1>Sunday, July 27th.</h1>
+
   <h1>Thursday, July 27th.</h1>
 +
<li> Cara Jones</li>
 +
<ul>Dilution of Mamba biobrick prefix/suffix, PCR of mamba in pGEX, PCR purification: <a href="https://static.igem.org/mediawiki/2017/7/7e/T--Georgia_State--7_27_17_cj.pdf"> See attached procedure for instructions.</a>  
 
</div>
 
</div>
  
 
<div id="date-7-28">
 
<div id="date-7-28">
−
   <h1>Monday, July 28th.</h1>
+
   <h1>Friday, July 28th.</h1>
 
</div>
 
</div>
  
 
<div id="date-7-29">
 
<div id="date-7-29">
−
   <h1>Tuesday, July 29th.</h1>
+
   <h1>Saturday, July 29th.</h1>
 
</div>
 
</div>
  
 
<div id="date-7-30">
 
<div id="date-7-30">
−
   <h1>Wednesday, July 30th.</h1>
+
   <h1>Sunday, July 30th.</h1>
 
</div>
 
</div>
  
 
<div id="date-7-31">
 
<div id="date-7-31">
−
   <h1>Thursday, July 31st.</h1>
+
   <h1>Monday, July 31st.</h1>
 
</div>
 
</div>
  
−
<!-- Agosto -->
+
<!-- August -->
 
<div id="date-8-1">
 
<div id="date-8-1">
 
   <h1>Tuesday,August 1st.</h1>
 
   <h1>Tuesday,August 1st.</h1>
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<ul><b>Results:</b><table class="w3-table"><tr><td><img src="https://static.igem.org/mediawiki/2017/6/62/T--Georgia_State--8.3DoubleRestrictionDigestHCG.jpg"></td><td>Since the diagnostic gel was a success the PSB1C backbone was extracted from the Lt 10 plasmid using an E-gel.</td></tr></table> </ul>
 
<ul><b>Results:</b><table class="w3-table"><tr><td><img src="https://static.igem.org/mediawiki/2017/6/62/T--Georgia_State--8.3DoubleRestrictionDigestHCG.jpg"></td><td>Since the diagnostic gel was a success the PSB1C backbone was extracted from the Lt 10 plasmid using an E-gel.</td></tr></table> </ul>
 
</div>
 
</div>
 +
 
<div id="date-8-6">
 
<div id="date-8-6">
 
   <h1>Sunday, August 6th.</h1>
 
   <h1>Sunday, August 6th.</h1>
 
</div>
 
</div>
 +
 +
  
 
<div id="date-8-7">
 
<div id="date-8-7">
 
   <h1>Monday, August 7th.</h1>
 
   <h1>Monday, August 7th.</h1>
 +
<li> Cara Jones</li>
 +
<ul><b>Objective:</b>Culture of transformed E.coli with mamba in PGEX</ul>
 +
<ul>I picked 10 colonies from plates and did overnight cultures in LB</ul>
 
</div>
 
</div>
  
 
<div id="date-8-8">
 
<div id="date-8-8">
 
   <h1>Tuesday, August 8th.</h1>
 
   <h1>Tuesday, August 8th.</h1>
 +
<li> Eudoxie Bataba</li>
 +
<ul><b>Objective:</b>I will get the mamba in PGEX plasmid from transformed E.coli</ul>
 +
<ul><b>Materials:</b>NEB miniprep kit, 50% glycerol</ul>
 +
<ul><b>Equipment:</b>Centrifuge and Nanophotometer</ul>
 +
<ul><b>Procedure:</b><a href="https://static.igem.org/mediawiki/2017/f/ff/T--Georgia_State--mamba_miniprep.jpg">See attached procedure.</a></ul>
 +
<ul><b>Results:</b> <table class="w3-table">
 +
<tr>
 +
<th>Sample</th>
 +
<th>Concentration (ng/uL)</th>
 +
</tr>
 +
<tr>
 +
<td>1</td>
 +
<td>118</td>
 +
</tr>
 +
<tr>
 +
<td>2</td>
 +
<td> 123</td>
 +
</tr>
 +
<tr>
 +
<td>3</td>
 +
<td> 85</td>
 +
</tr>
 +
<tr>
 +
<td>4</td>
 +
<td>133</td>
 +
</tr>
 +
<tr>
 +
<td>5</td>
 +
<td>90</td>
 +
</tr>
 +
<tr>
 +
<td>6</td>
 +
<td> 75</td>
 +
</tr>
 +
<tr>
 +
<td>7</td>
 +
<td> 72</td>
 +
</tr>
 +
<tr>
 +
<td>8</td>
 +
<td> 100</td>
 +
</tr>
 +
<tr>
 +
<td>9</td>
 +
<td> 65</td>
 +
</tr>
 +
<tr>
 +
<td>10</td>
 +
<td> 140</td>
 +
</tr>
 +
</table></ul>
 +
<br>
 +
<li> Cara Jones</li>
 +
<ul>SDS PAGE and Coomassie gel stain of purified mamba in pGEX: <a href="https://static.igem.org/mediawiki/2017/2/28/T--Georgia_State--8_8_17_cj.pdf"> See attached procedure for instructions.</a>
 
  </div>
 
  </div>
  
Line 660: Line 553:
 
</td></tr></table>
 
</td></tr></table>
 
  </ul>
 
  </ul>
 +
<li> Cara Jones</li>
 +
<ul>Re-do Restrction Digest of mamba: <a href="https://static.igem.org/mediawiki/2017/b/bd/T--Georgia_State--8_21_17_cj.pdf"> See attached procedure for instructions.</a>
  
 
</div>
 
</div>
Line 669: Line 564:
 
<ul><b>Objective:</b>To combine the four PCR-ed g-blocks of factor-C.</ul>
 
<ul><b>Objective:</b>To combine the four PCR-ed g-blocks of factor-C.</ul>
 
<ul><b>Procedure:</b> <a href="https://static.igem.org/mediawiki/2017/1/13/T--Georgia_State--OverlapPCRprotocol.pdf">See attached protocol for details</a></ul>
 
<ul><b>Procedure:</b> <a href="https://static.igem.org/mediawiki/2017/1/13/T--Georgia_State--OverlapPCRprotocol.pdf">See attached protocol for details</a></ul>
−
<ul><b>Results:</b><table class="w3-table"><tr><td><img src="https://static.igem.org/mediawiki/2017/0/08/T--Georgia_State--8.15SimulatedgelofOVERLAP.jpg"></td><td><img src="https://static.igem.org/mediawiki/2017/c/ca/T--Georgia_State--8.15gelofOVERLAP.jpg"></td></tr><tr><td></td><td></td></tr></ul>
+
<ul><b>Results:</b><table class="w3-table"><tr><td><img src="https://static.igem.org/mediawiki/2017/0/08/T--Georgia_State--8.15SimulatedgelofOVERLAP.jpg"></td><td><img src="https://static.igem.org/mediawiki/2017/c/ca/T--Georgia_State--8.15gelofOVERLAP.jpg"></td></tr><tr><td>Simulated gel</td><td>Wells contain the following:
 +
Gene Ruler 1kb plus MW ladder, Reaction of blocks 1 and 2, Negative control (blocks 1 and 2), Reaction of blocks 3 and 4, Negative control (blocks 3 and 4), Reaction of 1 and 2 with 3 and 4 products, negative control (reactions 1 and 2 with 3 and 3 and 4), and Gene Ruler 1kb plus MW ladder.
 +
</td></tr></table></ul>
 
</div>
 
</div>
  
Line 684: Line 581:
 
<ul><b>Equipment:</b>Thermocycler</ul>
 
<ul><b>Equipment:</b>Thermocycler</ul>
 
<ul><b>Procedure:</b>Two 50 ul PCR reactions were performed. 25ul of qiagen mastermix, 2ul of block 5 DNA, 2.5ul of forward primer, 2.5 ul of reverse primer and 18 ul of nuclease-free water was added to tube 1 and tube 2. </ul>  
 
<ul><b>Procedure:</b>Two 50 ul PCR reactions were performed. 25ul of qiagen mastermix, 2ul of block 5 DNA, 2.5ul of forward primer, 2.5 ul of reverse primer and 18 ul of nuclease-free water was added to tube 1 and tube 2. </ul>  
 +
<li> Tatenda Tela</li>
 +
<ul> Overnight cultures</ul>
 +
<ul> Colony PCR of Cara's transformants</ul>
 
</div>
 
</div>
  
Line 702: Line 602:
 
<div id="date-8-21">
 
<div id="date-8-21">
 
   <h1>Monday, August 21st.</h1>
 
   <h1>Monday, August 21st.</h1>
 +
<li>Tatenda Tela</li>
 +
<ul> Miniprep</ul>
 +
<ul>Ligations</ul>
 +
 
  </div>
 
  </div>
  
Line 711: Line 615:
 
<div id="date-8-23">
 
<div id="date-8-23">
 
   <h1>Wednesday, August 23rd.</h1>
 
   <h1>Wednesday, August 23rd.</h1>
 +
<li> Cara Jones</li>
 +
<ul>Diagnostic digest and electrophoresis of mamba: <a href="https://static.igem.org/mediawiki/2017/0/0b/T--Georgia_State--8_23_17_cj.pdf"> See attached procedure for instructions.</a>
 
    
 
    
 
</div>
 
</div>
Line 716: Line 622:
 
<div id="date-8-24">
 
<div id="date-8-24">
 
   <h1>Thursday, August 24th.</h1>
 
   <h1>Thursday, August 24th.</h1>
 +
<li> Cara Jones</li>
 +
<ul>Site directed mutagenesis of mambalgin: <a href="https://static.igem.org/mediawiki/2017/6/62/T--Georgia_State--8_24_17_cj.pdf"> See attached procedure for instructions.</a>
 
</div>
 
</div>
  
Line 726: Line 634:
 
<ul><b>Equipment:</b>Thermocycler</ul>
 
<ul><b>Equipment:</b>Thermocycler</ul>
 
<ul><b>Procedure:</b><a href="https://static.igem.org/mediawiki/2017/0/02/T--Georgia_State--hCGdigestprotocol.pdf">See attached procedure.</a></ul>  
 
<ul><b>Procedure:</b><a href="https://static.igem.org/mediawiki/2017/0/02/T--Georgia_State--hCGdigestprotocol.pdf">See attached procedure.</a></ul>  
 +
<li> Cara Jones</li>
 +
<ul>Diagnostic gel of mutanagized mamba: <a href="https://static.igem.org/mediawiki/2017/5/54/T--Georgia_State--8_25_17_cj.pdf"> See attached procedure for instructions.</a>
 
</div>
 
</div>
  
Line 738: Line 648:
 
<div id="date-8-28">
 
<div id="date-8-28">
 
   <h1>Monday, August 28th.</h1>
 
   <h1>Monday, August 28th.</h1>
 +
<li> Cara Jones</li>
 +
<ul>Transformation of Mutagenesis RxNs: <a href="https://static.igem.org/mediawiki/2017/4/4e/T--Georgia_State--8_28_17_cj.pdf"> See attached procedure for instructions.</a>
 
   </div>
 
   </div>
  
Line 761: Line 673:
 
<div id="date-8-30">
 
<div id="date-8-30">
 
   <h1>Wednesday, August 30th.</h1>
 
   <h1>Wednesday, August 30th.</h1>
 +
<li> Cara Jones</li>
 +
<ul>Protein Expression, mamba in pGEX: <a href="https://static.igem.org/mediawiki/2017/e/e1/T--Georgia_State--8_30_17_cj.pdf"> See attached procedure for instructions.</a>
 
</div>
 
</div>
  
Line 770: Line 684:
 
<div id="date-9-1">
 
<div id="date-9-1">
 
   <h1>Friday, September 1st.</h1>
 
   <h1>Friday, September 1st.</h1>
−
   <ul>
+
    
−
    <li>Apoptosis kit arrived!, 14:00.</li>
+
−
    <li>Accutase arrived!, 14:00.</li>
+
−
    <li>Genes arrived!, 14:00.</li>
+
−
    <li>Preparation of LB and LB-agar media, 13:30.</li>
+
−
    <ul>
+
−
      <li>Eduardo Ramirez (1).</li>
+
−
    </ul>
+
−
    <li>Preparation of the plaques for the transformation, 17:00.</li>
+
−
    <ul>
+
−
      <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
  </ul>
+
−
  <table>
+
−
    <tbody>
+
−
      <tr>
+
−
        <td>
+
−
          <p>Abbreviation</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>Construct</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>Gene</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>Resistance</p>
+
−
        </td>
+
−
      </tr>
+
−
      <tr>
+
−
        <td>
+
−
          <p>LPP</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>C1</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>LPP Red Lambda</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>Amp</p>
+
−
        </td>
+
−
      </tr>
+
−
      <tr>
+
−
        <td>
+
−
          <p>MSBB</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>C2</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>Red Lambda</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>Kn + Amp</p>
+
−
        </td>
+
−
      </tr>
+
−
      <tr>
+
−
        <td>
+
−
          <p>H-GFP</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>C4</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>GFP</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>Amp</p>
+
−
        </td>
+
−
      </tr>
+
−
      <tr>
+
−
        <td>
+
−
          <p>C-Apop</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>C5</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>Apoptine</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>Amp</p>
+
−
        </td>
+
−
      </tr>
+
−
      <tr>
+
−
        <td>
+
−
          <p>H-SiRNA 1</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>C6</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>SiRNA 1</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>Amp</p>
+
−
        </td>
+
−
      </tr>
+
−
      <tr>
+
−
        <td>
+
−
          <p>H-SiRNA 2</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>C7</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>SiRNA 2</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>Amp</p>
+
−
        </td>
+
−
      </tr>
+
−
      <tr>
+
−
        <td>
+
−
          <p>H-SiRNA 3</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>C8</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>SiRNA 3</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>Amp</p>
+
−
        </td>
+
−
      </tr>
+
−
      <tr>
+
−
        <td>
+
−
          <p>H-SiRNA-Apop</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>C9</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>SiRNA + Apoptine</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>Amp</p>
+
−
        </td>
+
−
      </tr>
+
−
      <tr>
+
−
        <td>
+
−
          <p>BBa_1</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>Biobrick</p>
+
−
        </td>
+
−
        <td>&nbsp;</td>
+
−
        <td>
+
−
          <p>Cam</p>
+
−
        </td>
+
−
      </tr>
+
−
      <tr>
+
−
        <td>
+
−
          <p>BBa_2</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>Biobrick</p>
+
−
        </td>
+
−
        <td>&nbsp;</td>
+
−
        <td>
+
−
          <p>Cam</p>
+
−
        </td>
+
−
      </tr>
+
−
      <tr>
+
−
        <td>
+
−
          <p>BBa_3</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>Biobrick</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>Peptide + p3</p>
+
−
        </td>
+
−
        <td>
+
−
          <p>Cam</p>
+
−
        </td>
+
−
      </tr>
+
−
    </tbody>
+
−
  </table>
+
−
  <ul>
+
−
    <li>HyperLadder 50bp arrived!, 17:10</li>
+
−
    <li>HLA arrived!, 17:10:</li>
+
−
    <li>Transformation of C1 through C7 in TOP10, 19:00.</li>
+
−
    <ul>
+
−
      <li>Minerva.</li>
+
−
    </ul>
+
−
    <li>Transformation of GFP and RFP in TOP10, 21:30.</li>
+
−
    <ul>
+
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
  </ul>
+
 
</div>
 
</div>
  
 
<div id="date-9-2">
 
<div id="date-9-2">
−
   <h1>Tuesday, September 2</h1>
+
   <h1>Saturday, September 2nd.</h1>
−
   <ul>
+
    
−
    <li>Inoculation of plates with transformed colonies from yesterday, 12:00.</li>
+
−
    <ul>
+
−
      <li>Minerva.</li>
+
−
    </ul>
+
−
    <li>Transformations of GFP Amp, RFP Cam, D1 Kn, D2 Cam, D3 Amp, 19:30.</li>
+
−
    <ul>
+
−
      <li>Eduardo Ramirez (2).</li>
+
−
    </ul>
+
−
    <li>Miniprep of the grown transformed cells from yesterday (C4, C5, C6, C7, D2, GFP, D1, BBa_1), 21:00.</li>
+
−
    <ul>
+
−
      <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
  </ul>
+
 
</div>
 
</div>
  
 
<div id="date-9-3">
 
<div id="date-9-3">
−
   <h1>Wednesday, September 3</h1>
+
   <h1>Sunday, September 3rd</h1>
−
   <ul>
+
    
−
    <li>Digestions of the last minipreps, 02:00.</li>
+
−
    <ul>
+
−
      <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
    <li>Electrophoresis, 03:30.</li>
+
−
    <ul>
+
−
      <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
    <li>Preparation of electro-competent TOP10 cells, 08:00.</li>
+
−
    <ul>
+
−
      <li>Newbies: Alex, Danny, Chuy, Caro.</li>
+
−
    </ul>
+
−
    <li>Preparation of stock cells (C4, C5, C6, C7, D2, GFP, D1, BBa_1), 18:30.</li>
+
−
    <ul>
+
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
    <li>Ligations of the C4, C5, C6, C7 with the pcb1c3, 20:30.</li>
+
−
    <ul>
+
−
      <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
    <li>Inoculation of TOP10 for electro-competent cells tomorrow and D2 cells, 01:25.</li>
+
−
    <ul>
+
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
  </ul>
+
 
</div>
 
</div>
  
 
<div id="date-9-4">
 
<div id="date-9-4">
−
   <h1>Thursday, September 4</h1>
+
   <h1>Monday, September 4th</h1>
 
</div>
 
</div>
  
 
<div id="date-9-5">
 
<div id="date-9-5">
−
   <h1>Friday, September 5</h1>
+
   <h1>Tuesday, September 5th</h1>
−
   <ul>
+
    
−
    <li>Preparation of the tickets for the cake raffle (fundraising), 12:00.</li>
+
−
    <ul>
+
−
      <li>Cinthya, Omar, Eduardo Ramirez (1), Eduardo Cepeda, Mercedes.</li>
+
−
    </ul>
+
−
    <li>Miniprep, 14:00.</li>
+
−
    <ul>
+
−
      <li>Mercedes and Eduardo Cepeda with special guest: Ana Cristina.</li>
+
−
    </ul>
+
−
    <li>Talk with Professor Cardineau about the legislation proposal, 17:00.</li>
+
−
    <ul>
+
−
      <li>Omar.</li>
+
−
    </ul>
+
−
    <li>Preparations of electro-competente TOP10 cells, 17:30.</li>
+
−
    <ul>
+
−
      <li>Eduardo Ramirez (2), Mercedes.</li>
+
−
    </ul>
+
−
    <li>Digestions, 19:00.</li>
+
−
    <ul>
+
−
      <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
    <li>Inoculation of LB-agar plaques, 21:00.</li>
+
−
    <ul>
+
−
      <li>Cynthia.</li>
+
−
    </ul>
+
−
  </ul>
+
 
</div>
 
</div>
  
 
<div id="date-9-6">
 
<div id="date-9-6">
−
   <h1>Saturday, September 6</h1>
+
   <h1>Wednesday, September 6</h1>
 
</div>
 
</div>
  
 
<div id="date-9-7">
 
<div id="date-9-7">
−
   <h1>Sunday, September 7</h1>
+
   <h1>Thursday, September 7</h1>
 
</div>
 
</div>
  
 
<div id="date-9-8">
 
<div id="date-9-8">
−
   <h1>Monday, September 8</h1>
+
   <h1>Friday, September 8th.</h1>
 +
<li> Tatenda Tela</li>
 +
<ul>hCG for pGEX pcr</ul>
 +
<ul><b>Results</b>:<table><tr><td><img src="https://static.igem.org/mediawiki/2017/f/fb/T--Georgia_State--091317_PCR_of_HCG-Tatenda.jpeg" alt=""></td></table>
 +
<table><tr><th>lane</th><th>contents</th></tr>
 +
<tr><td>1</td><td>Generuler 1kb plus ladder</td></tr>
 +
<tr><td>2</td><td>hCG insert w/ hCG block 5 fwd&rev primers</td></tr>
 +
<tr><td>3</td><td>hCG insert w/ hCG block 5 fwd&rev primers</td></tr>
 +
<tr><td>4</td><td>hCG insert w/ hCG block 5 fwd&rev primers</td></tr>
 +
<tr><td>5</td><td>hCG insert w/ hCG block 5 fwd&rev primers</td></tr>
 +
<tr><td>6</td><td>hCG insert w/ hCG block 5 fwd&rev primers</td></tr>
 +
<tr><td>7</td><td>hCG insert w/ hCG block 5 fwd&rev primers</td></tr>
 +
<tr><td>8</td><td>hCG insert w/ hCG for pGEX fwd&rev primers</td></tr>
 +
<tr><td>9</td><td>hCG insert w/ hCG for pGEX fwd&rev primers</td></tr>
 +
<tr><td>10</td><td>hCG insert w/ hCG for pGEX fwd&rev primers</td></tr>
 +
<tr><td>11</td><td>hCG insert w/ hCG for pGEX fwd&rev primers</td></tr>
 +
<tr><td>12</td><td>hCG insert w/ hCG for pGEX fwd&rev primers</td></tr>
 +
<tr><td>13</td><td>hCG insert w/ hCG for pGEX fwd&rev primers</td></tr>
 +
<tr><td>14</td><td>Generuler 1kb plus ladder</td></tr>
 +
 
 +
</table></ul>
 
</div>
 
</div>
  
 
<div id="date-9-9">
 
<div id="date-9-9">
−
   <h1>Tuesday, September 9</h1>
+
   <h1>Saturday, September 9</h1>
−
  <ul>
+
−
    <li>Inoculation of PCB1c3 (with C4, C5, C7 separately), 13:30.</li>
+
−
    <ul>
+
−
      <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
  </ul>
+
 
</div>
 
</div>
  
 
<div id="date-9-10">  
 
<div id="date-9-10">  
−
   <h1>Wednesday, September 10</h1>
+
   <h1>Sunday, September 10</h1>
 
</div>
 
</div>
  
 
<div id="date-9-11">  
 
<div id="date-9-11">  
−
   <h1>Thursday, September 11</h1>
+
   <h1>Monday, September 11</h1>
−
  <ul>
+
−
    <li>Revision of (not grown yet) plates, 12:00.</li>
+
−
    <ul>
+
−
      <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
    <li>Inoculaton of D2-GFP, 21:00.</li>
+
−
    <ul>
+
−
      <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
  </ul>
+
 
</div>
 
</div>
  
 
<div id="date-9-12">
 
<div id="date-9-12">
−
   <h1>Friday, September 12</strong><strong>th</strong></h1>
+
   <h1>Tuesday, September 12</h1>
−
  <ul>
+
−
    <li>Cleaning of the electroporation cells for the electro-competent cells, 16:00.</li>
+
−
    <ul>
+
−
      <li>Minerva.</li>
+
−
    </ul>
+
−
    <li>Inoculation of cells that grew from the prior day, 18:00.</li>
+
−
    <ul>
+
−
      <li>Cinthya.</li>
+
−
    </ul>
+
−
  </ul>
+
 
</div>
 
</div>
  
 
<div id="date-9-13">  
 
<div id="date-9-13">  
−
   <h1>Saturday, September 13</h1>
+
   <h1>Wednesday, September 13</h1>
−
  <ul>
+
<li>Cara Jones</li>
−
    <li>Transformation of the D2 and D3 variants for the interlab project, 00:10.</li>
+
<ul>Overnight Cultures</ul>
−
    <ul>
+
<li> Tatenda Tela</li>
−
      <li>Eduardo Ramirez (2), Mercedes, Claudia, Omar.</li>
+
<ul>Restriction digest and heat inactivation of samples from September 8</ul>
−
    </ul>
+
 
−
  </ul>
+
 
</div>
 
</div>
  
 
<div id="date-9-14">  
 
<div id="date-9-14">  
−
   <h1>Sunday, September 14th.</strong></h1>
+
   <h1>Thursday, September 14th.</h1>
−
  <ul>
+
<li>Tatenda Tela</li>
−
    <li>Miniprep, 17:00.</li>
+
<ul>Miniprep of overnight cultures prepped by Cara the day before</ul>
−
    <ul>
+
<ul><b>Results</b>:<table class="w3-table">
−
      <li>Mercedes, Eduardo Ramirez (2)</li>
+
<tr>
−
    </ul>
+
<th>Sample</th>
−
    <li>Sterilization of Falcon tubes, LB broth and plastic pearls, 18:30.</li>
+
<th>Concentration (ng/uL)</th>
−
    <ul>
+
<th>A260/A280</th>
−
      <li>Omar</li>
+
</tr>
−
    </ul>
+
<tr>
−
    <li>Digestions, 21:00.</li>
+
<td>1</td>
−
    <ul>
+
<td>25</td><td>2.0</td>
−
      <li>Mercedes.</li>
+
</tr>
−
    </ul>
+
<tr>
−
    <li>Inoculation of the transformed cells with D2 and D3 variants, 22:00.</li>
+
<td>2</td>
−
    <ul>
+
<td>7.5</td><td>2.0</td>
−
      <li>Omar.</li>
+
</tr>
−
    </ul>
+
<tr>
−
  </ul>
+
<td>3</td>
 +
<td>9.8</td>
 +
<td>1.7</td>
 +
</tr>
 +
<tr>
 +
<td>4</td>
 +
<td>8.5</td>
 +
<td>1.8</td>
 +
</tr>
 +
<tr>
 +
<td>5</td>
 +
<td>9.5</td>
 +
<td>1.7</td>
 +
</tr>
 +
</table></ul>
 +
<br>
 +
<li> Cara Jones</li>
 +
<ul>Interlab: Media Preparation: <a href="https://static.igem.org/mediawiki/2017/3/3d/T--Georgia_State--9_14_17_cj.pdf"> See attached procedure for instructions.</a>  
 
</div>
 
</div>
 +
  
 
<div id="date-9-15">  
 
<div id="date-9-15">  
−
   <h1>Monday, September 15</h1>
+
   <h1>Friday, September 15</h1>
−
  <ul>
+
−
    <li>Miniprep of yesterday's inoculations of D2 and D3 variants, 10:00.</li>
+
−
    <ul>
+
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
    <li>Digestions, 17:30.</li>
+
−
    <ul>
+
−
      <li>Eduardo Ramirez (2).</li>
+
−
    </ul>
+
−
    <li>Inoculation of TOP10 and XL-Blue (Tet), 22:50.</li>
+
−
    <ul>
+
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
  </ul>
+
 
</div>   
 
</div>   
  
 
  <div id="date-9-16">   
 
  <div id="date-9-16">   
−
   <h1>Tuesday, September 16th.</h1>
+
   <h1>Saturday, September 16th.</h1>
−
  <ul>
+
<li>Victoria Radcliffe, Aditya Natu</li>
−
    <li>Transformations of the M13 bacteriophage into the XL-Blue, 10:00.</li>
+
<ul>Midiprep of mamba in pGEX to get pGEx backbone</ul>  
−
    <ul>
+
<table><tr><th>sample</th><th>concentration (ng/uL)</th></tr>
−
      <li>Mercedes.</li>
+
<tr><td>1</td><td>200</td></tr>
−
    </ul>
+
<tr><td>2</td><td>275</td></tr>
−
  </ul>
+
</table>
 
</div>
 
</div>
  
 
<div id="date-9-17">  
 
<div id="date-9-17">  
−
   <h1>Wednesday, September 17</h1>
+
   <h1>Sunday, September 17</h1>
−
  <ul>
+
−
    <li>Miniprep 01:15.</li>
+
−
    <ul>
+
−
      <li>Minerva, Eduardo Cepeda.</li>
+
−
    </ul>
+
−
    <li>Digestions (C4, C5, C6, C7 and , 02:53.</li>
+
−
    <ul>
+
−
      <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
    <li>PBS buffer, 12:30.</li>
+
−
    <ul>
+
−
      <li>Omar.</li>
+
−
    </ul>
+
−
    <li>Recovery of DNA from Gel, 13:30.</li>
+
−
    <ul>
+
−
      <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
  </ul>
+
 
</div>
 
</div>
  
 
<div id="date-9-18">  
 
<div id="date-9-18">  
−
   <h1>Thursday, September 18</h1>
+
   <h1>Monday, September 18</h1>
−
  <ul>
+
−
    <li>Transformations of C8, C9 and ligation in TOP10 electro-competent, 00:40.</li>
+
−
    <ul>
+
−
      <li>Cinthya, Omar.</li>
+
−
    </ul>
+
−
    <li>Electrophoresis, 01:30.</li>
+
−
    <ul>
+
−
      <li>Mercedes, Eduardo Ramirez (1).</li>
+
−
    </ul>
+
−
    <li>Plates from the transformations, 01:50.</li>
+
−
    <ul>
+
−
      <li>Omar.</li>
+
−
    </ul>
+
−
    <li>Miniprep of PET, 12:30.</li>
+
−
    <ul>
+
−
      <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
    <li>Transformation replica of the same ligation as before, 15:30.</li>
+
−
    <ul>
+
−
      <li>Omar.</li>
+
−
    </ul>
+
−
    <li>Preparation of LB broth, 17:00.</li>
+
−
    <ul>
+
−
      <li>All the newbies (Caro, Jesus, Alex, Dany).</li>
+
−
    </ul>
+
−
    <li>Digestion of PET, 19:30.</li>
+
−
    <ul>
+
−
      <li>Claudia.</li>
+
−
    </ul>
+
−
  </ul>
+
 
</div>
 
</div>
  
 
<div id="date-9-19">  
 
<div id="date-9-19">  
−
   <h1>Friday, September 19</h1>
+
   <h1>Tuesday, September 19</h1>
−
  <ul>
+
<li> Tatenda Tela</li>
−
    <li>Ligation of PET with all the constructs, 01:19.</li>
+
<ul> Digest of Midiprep of mamba in pGEX done by Victoria and Aditya using EcoRI and NotI enzymes</ul>
−
    <ul>
+
<ul><b>Results</b>:<table><tr><td><img src="https://static.igem.org/mediawiki/2017/9/9c/T--Georgia_State--091917_Restriction_digest_of_hcg_in_pGEX-Tatenda.jpeg" alt=""></td></table>
−
      <li>Eduardo Cepeda.</li>
+
<table><tr><th>lane</th><th>contents</th></tr>
−
    </ul>
+
<tr><td>1</td><td>Generuler 1kb plus ladder</td></tr>
−
    <li>Electrophoresis, 03:30.</li>
+
<tr><td>2</td><td>Mamba in pGEX</td></tr>
−
    <ul>
+
<tr><td>3</td><td>Mamba in pGEX</td></tr>
−
      <li>Eduardo Cepeda.</li>
+
<tr><td>4</td><td>Mamba in pGEX cut with EcoRI</td></tr>
−
    </ul>
+
<tr><td>5</td><td>Mamba in pGEX cut with EcoRI</td></tr>
−
    <li>Miniprep, 8:30.</li>
+
<tr><td>6</td><td>empty</td></tr>
−
    <ul>
+
<tr><td>7</td><td>Mamba in pGEX cut with EcoRI and NotI</td></tr>
−
      <li>Mercedes.</li>
+
<tr><td>8</td><td>empty</td></tr>
−
    </ul>
+
<tr><td>9</td><td>Mamba in pGEX cut with EcoRI and NotI</td></tr>
−
    <li>Digestions, 13:00.</li>
+
<tr><td>10</td><td>empty</td></tr>
−
    <ul>
+
<tr><td>11</td><td>Mamba in pGEX cut with EcoRI and PstI</td></tr>
−
      <li>Mercedes.</li>
+
<tr><td>12</td><td>Mamba in pGEX cut with EcoRI and PstI</td></tr>
−
    </ul>
+
<tr><td>13</td><td>empty</td></tr>
−
    <li>Electrophoresis, 15:30.</li>
+
<tr><td>14</td><td>empty</td></tr></table></ul>
−
    <ul>
+
<li> Tatenda Tela</li>
−
      <li>Mercedes.</li>
+
<ul> Gel extraction of pGEX backbone (lanes 7 and 9)</ul>
−
    </ul>
+
<ul> Ligation of digested pGEX backbone and digested hCG from September 13</ul>
−
    <li>Quantification of the fluorescence for the interlab project, 16:30.</li>
+
<ul> Transforamtion using 5 alpha competent cells
−
    <ul>
+
−
      <li>Mercedes, Eduardo Ramirez (2).</li>
+
−
    </ul>
+
−
    <li>Group photo taken, 18:00.</li>
+
−
    <ul>
+
−
      <li>Everyone except Mercedes, Eduardo Cepeda and Eduardo Ramirez.</li>
+
−
    </ul>
+
−
    <li>Group video filmed, 18:30.</li>
+
−
    <ul>
+
−
      <li>Everyone.</li>
+
−
    </ul>
+
−
    <li>Transformation of TOP10 cells with PET + (C4, C5, C6, C7), 20:30.</li>
+
−
    <ul>
+
−
      <li>Cinthya, Eduardo Cepeda.</li>
+
−
    </ul>
+
−
    <li>Transformation of TOP10 cells with various repetitions of D2 and D3 (with GFP or RFP), 21:00.</li>
+
−
    <ul>
+
−
      <li>Mercedes, Eduardo Ramirez (2).</li>
+
−
    </ul>
+
−
    <li>Inoculation in Falcon tubes with the ligation of C5 and various others, 22:00.</li>
+
−
    <ul>
+
−
      <li>Omar.</li>
+
−
    </ul>
+
−
    <li>Inoculation in plates of the previous transformed TOP10 cells with PET, 23:00.</li>
+
−
    <ul>
+
−
      <li>Cinthya.</li>
+
−
    </ul>
+
−
  </ul>
+
 
</div>
 
</div>
  
 
<div id="date-9-20">  
 
<div id="date-9-20">  
−
   <h1>Saturday, September 20</h1>
+
   <h1>Wednesday, September 20</h1>
−
  <ul>
+
−
    <li>Transformation in electro-competent cells, 01:20.</li>
+
−
    <ul>
+
−
      <li>Mercedes, Eduardo Ramirez.</li>
+
−
    </ul>
+
−
    <li>Minipreps, 18:00.</li>
+
−
    <ul>
+
−
      <li>Mercedes, Omar.</li>
+
−
    </ul>
+
−
    <li>Digestions, 22:00.</li>
+
−
  </ul>
+
 
</div>
 
</div>
  
 
<div id="date-9-21">  
 
<div id="date-9-21">  
−
   <h1>Sunday, September 21</strong><strong>st</strong><strong>.</strong></h1>
+
   <h1>Thursday, September 21</h1>
−
  <ul>
+
<li>Tatenda Tela</li>
−
    <li>Electrophoresis, 00:00.</li>
+
<ul>Another digest mamba in pGEX and then a ligation with the hCG insert followed by a transformation into 5 alpha competent cells</ul>
−
    <ul>
+
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
    <li>Inoculations for minipreps, 01:00.</li>
+
−
    <ul>
+
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
    <li>Minipreps,</li>
+
−
    <ul>
+
−
      <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
  </ul>
+
 
</div>
 
</div>
  
 
<div id="date-9-22">  
 
<div id="date-9-22">  
−
   <h1>Monday, September 22</h1>
+
   <h1>Friday, September 22</h1>
−
  <ul>
+
<li>Julia Ngo<li>
−
    <li>Miniprep, 08:00.</li>
+
<ul> PCR of hCG using hCG for pGEX fwd and rev primers</ul>
−
    <ul>
+
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
    <li>Revision of the cell stocks, contamination detected, 09:00.</li>
+
−
    <ul>
+
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
    <li>Inoculations of C8 and C9, 22:00.</li>
+
−
    <ul>
+
−
      <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
  </ul>
+
 
</div>
 
</div>
  
 
<div id="date-9-23">  
 
<div id="date-9-23">  
−
   <h1>Tuesday, September 23</h1>
+
   <h1>Saturday, September 23</h1>
−
  <ul>
+
−
    <li>Preparation of electo-competent cells DH5a, 03:00.</li>
+
−
    <ul>
+
−
      <li>Minerva.</li>
+
−
    </ul>
+
−
    <li>Minipreps for the interlab, 06:00</li>
+
−
    <ul>
+
−
      <li>Mercedes, Minerva.</li>
+
−
    </ul>
+
−
    <li>Transformations for the interlab, 07:30.</li>
+
−
    <ul>
+
−
      <li>Mercedes, Minerva.</li>
+
−
    </ul>
+
−
    <li>Digestions for the interlab, 12:30.</li>
+
−
    <ul>
+
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
    <li>Miniprep, 13:30.</li>
+
−
    <ul>
+
−
      <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
    <li>Electrophoresis for interlab, 19:00.</li>
+
−
    <ul>
+
−
      <li>Eduardo Ramirez (2)</li>
+
−
    </ul>
+
−
    <li>Electrophoresis for the project (C8, C9), 21:00.</li>
+
−
    <ul>
+
−
      <li>Eduardo Ramirez (2)</li>
+
−
    </ul>
+
−
  </ul>
+
 
</div>
 
</div>
  
 
<div id="date-9-24">
 
<div id="date-9-24">
−
   <h1>Wednesday, September 24</h1>
+
   <h1>Sunday, September 24</h1>
−
  <ul>
+
−
    <li>Purification of the electrophoresis gel, 01:20.</li>
+
−
    <ul>
+
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
    <li>Digestions for the interlab, 01:20.</li>
+
−
    <ul>
+
−
      <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
    <li>Ligations, 02:00.</li>
+
−
    <ul>
+
−
      <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
    <li>Electrophoresis for the interlab, 02:40.</li>
+
−
    <ul>
+
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
    <li>Inoculations for the interlab, 11:00.</li>
+
−
    <ul>
+
−
      <li>Omar, Mercedes.</li>
+
−
    </ul>
+
−
  </ul>
+
 
</div>
 
</div>
  
 
<div id="date-9-25">  
 
<div id="date-9-25">  
−
   <h1>Thursday, September 25</h1>
+
   <h1>Monday, September 25</h1>
−
  <ul>
+
<li>Julia Ngo</li>
−
    <li>Inoculation of XL-Blue, 22:00.</li>
+
<ul>Ran gel of hCG PCR from September 22</ul>
−
    <ul>
+
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
    <li>Infection of the XL-Blue with bacteriophages, 23:00.</li>
+
−
    <ul>
+
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
  </ul>
+
 
</div>
 
</div>
  
 
<div id="date-9-26">   
 
<div id="date-9-26">   
−
   <h1>Friday, September 26</h1>
+
   <h1>Tuesday, September 26</h1>
−
  <ul>
+
−
    <li>Miniprep of the bacteriophage-infected cells, 03:00.</li>
+
−
    <ul>
+
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
    <li>Preparation of the cells for quantification of fluorescence and OD for the interlab project, from 8:00 to 24:00.</li>
+
−
    <ul>
+
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
    <li>Purification of the miniprep. 18:00.</li>
+
−
    <ul>
+
−
      <li>Eduardo Ramirez (1), Claudia.</li>
+
−
    </ul>
+
−
    <li>Transformations of C1, C2, Bba_1, Bba_2, and ligations, 19:00.</li>
+
−
    <ul>
+
−
      <li>Cinthya, Omar.</li>
+
−
    </ul>
+
−
    <li>Plating of the transformations, 22:00.</li>
+
−
    <ul>
+
−
      <li>Cinthya.</li>
+
−
    </ul>
+
−
  </ul>
+
 
</div>
 
</div>
  
 
<div id="date-9-27">  
 
<div id="date-9-27">  
−
   <h1>Saturday, September 27</h1>
+
   <h1>Wednesday, September 27</h1>
−
  <ul>
+
</div>
−
    <li>Inoculation of C1, C2, Bba_1, Bba _2 and ligations, 20:00.</li>
+
−
    <ul>
+
−
      <li>Omar</li>
+
−
    </ul>
+
−
    <li>Sterilization of material, 22:00.</li>
+
−
    <ul>
+
−
      <li>Omar.</li>
+
−
    </ul>
+
−
  </ul>
+
−
</div>
+
  
 
<div id="date-9-28">  
 
<div id="date-9-28">  
−
   <h1>Sunday, September 28</h1>
+
   <h1>Thursday, September 28</h1>
−
  <ul>
+
−
    <li>Preparation of electro-competent cells, 01:00.</li>
+
−
    <ul>
+
−
      <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
    <li>Ligations, 03:00.</li>
+
−
    <ul>
+
−
      <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
    <li>Electrophoresis, 12:00.</li>
+
−
    <ul>
+
−
      <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
    <li>Minipreps of the inoculations from Saturday, 13:00.</li>
+
−
    <ul>
+
−
      <li>Claudia, Eduardo Ramirez (1).</li>
+
−
    </ul>
+
−
    <li>Recovery of DNA from gel, 15:00.</li>
+
−
    <ul>
+
−
      <li>Claudia.</li>
+
−
    </ul>
+
−
  </ul>
+
 
</div>
 
</div>
  
 
<div id="date-9-29">  
 
<div id="date-9-29">  
−
   <h1>Monday, September 29</h1>
+
   <h1>Friday, September 29</h1>
 +
<li>Julia Ngo</li>
 +
<ul>Designing Primers for Tac-GST-Mamba in pSB1C3</ul>
 
</div>
 
</div>
  
 
<div id="date-9-30">  
 
<div id="date-9-30">  
−
   <h1>Tuesday, September 30</h1>
+
   <h1>Saturday, September 30</h1>
 
</div>
 
</div>
 +
  
 
<!-- October -->
 
<!-- October -->
 
<div id="date-10-1">
 
<div id="date-10-1">
−
   <h1>Wednesday, October 1st.</h1>
+
   <h1>Sunday, October 1st.</h1>
−
  <ul>
+
−
    <li>Minipreps of all constructs, 12:00.</li>
+
−
    <ul>
+
−
      <li>Eduardo.</li>
+
−
    </ul>
+
−
    <li>Digestions of all constructs, 20:30.</li>
+
−
    <ul>
+
−
      <li>Claudia, Jesus.</li>
+
−
    </ul>
+
−
    <li>Miniprep of C2, 21:00.</li>
+
−
    <ul>
+
−
      <li>Jesus.</li>
+
−
    </ul>
+
−
  </ul>
+
 
</div>
 
</div>
  
 
<div id="date-10-2">
 
<div id="date-10-2">
−
   <h1>Thursday, October 2nd.</h1>
+
   <h1>Monday, October 2nd.</h1>
−
  <ul>
+
<li> Cara Jones</li>
−
    <li>Collaboration: gave C4 through C9 to the UANL team, 13:30.</li>
+
<ul>Mamba Mutagenesis: <a href="https://static.igem.org/mediawiki/2017/5/5c/T--Georgia_State--10_2_17_cj.pdf"> See attached procedure for instructions.</a>  
−
    <ul>
+
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
    <li>Inoculo XL-Blue, 18:00.</li>
+
−
    <ul>
+
−
      <li>Eduardo Ramirez (2).</li>
+
−
    </ul>
+
−
    <li>Miniprep C8, C10, Bb_1, C6, 16:30.</li>
+
−
    <ul>
+
−
      <li>Eduardo Cepeda, Eduardo Ramirez (2).</li>
+
−
    </ul>
+
−
    <li>Miniprep of phagemides, 18:30.</li>
+
−
    <ul>
+
−
      <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
  </ul>
+
 
</div>
 
</div>
  
 
<div id="date-10-3">
 
<div id="date-10-3">
−
   <h1>Friday, October 3rd.</h1>
+
   <h1>Tuesday, October 3rd.</h1>
−
  <ul>
+
<li>Julia Ngo</li>
−
    <li>Digestions of C1, C2 and PET variants of the other constructs, 12:00.</li>
+
<ul>PCR of Tac promoter</ul>
−
    <ul>
+
−
      <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
    <li>Minipreps, 12:00.</li>
+
−
    <ul>
+
−
      <li>Claudia.</li>
+
−
    </ul>
+
−
    <li>Electrophoresis, 16:00.</li>
+
−
    <ul>
+
−
      <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
    <li>Inoculations, 20:00.</li>
+
−
    <ul>
+
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
  </ul>
+
 
</div>
 
</div>
  
 
<div id="date-10-4">
 
<div id="date-10-4">
−
   <h1>Saturday, October 4th.</h1>
+
   <h1>Wednesday, October 4th.</h1>
 +
<li> Cara Jones</li>
 +
<ul>Diagnostic Restriction Digest, Mutagenesis, Transformation: <a href="https://static.igem.org/mediawiki/2017/3/3e/T--Georgia_State--10_4_17_cj.pdf"> See attached procedure for instructions.</a>
 +
<li>Julia Ngo</li>
 +
<ul> PCR Cleanup</ul>
 +
<ul>Diagnostic Gel of PCR</ul>
 
</div>
 
</div>
  
 
<div id="date-10-5">
 
<div id="date-10-5">
−
   <h1>Sunday, October 5th.</h1>
+
   <h1>Thursday, October 5th.</h1>
−
  <ul>
+
−
    <li>Western Blot, 01:00.</li>
+
−
    <ul>
+
−
      <li>Minerva.</li>
+
−
    </ul>
+
−
    <li>Minipreps, 00:30.</li>
+
−
    <ul>
+
−
      <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
    <li>Electrophoresis, 04:00.</li>
+
−
    <ul>
+
−
      <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
    <li>Inoculations, 18:00.</li>
+
−
    <ul>
+
−
      <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
  </ul>
+
 
</div>
 
</div>
  
 
<div id="date-10-6">
 
<div id="date-10-6">
−
   <h1>Monday, October 6th.</h1>
+
   <h1>Friday, October 6th.</h1>
 
</div>
 
</div>
  
 
<div id="date-10-7">
 
<div id="date-10-7">
−
   <h1>Tuesday, October 7th.</h1>
+
   <h1>Saturday, October 7th.</h1>
 
</div>
 
</div>
  
 
<div id="date-10-8">
 
<div id="date-10-8">
−
   <h1>Wednesday, October 8th.</h1>
+
   <h1>Sunday, October 8th.</h1>
 
</div>
 
</div>
  
 
<div id="date-10-9">
 
<div id="date-10-9">
−
   <h1>Thursday, October 9th.</h1>
+
   <h1>Monday, October 9th.</h1>
−
  <ul>
+
<li> Cara Jones</li>
−
    <li>Minipreps for cells with flippase, 12:00.</li>
+
<ul>Colony PCR, Digest of PCR: <a href="https://static.igem.org/mediawiki/2017/e/ef/T--Georgia_State--10_9_17_cj.pdf"> See attached procedure for instructions.</a>  
−
    <ul>
+
<li> Julia Ngo</li>
−
      <li>Eduardo Cepeda.</li>
+
<ul>Redo of PCR of Tac Promoter</ul>
−
    </ul>
+
−
    <li>Stock of cells with flippase, 12:00.</li>
+
−
    <ul>
+
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
    <li>Digestions for flippase, 16:00.</li>
+
−
    <ul>
+
−
      <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
    <li>Electrophoresis, 18:00.</li>
+
−
    <ul>
+
−
      <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
  </ul>
+
 
</div>
 
</div>
  
 
<div id="date-10-10">
 
<div id="date-10-10">
−
   <h1>Friday, October 10th.</h1>
+
   <h1>Tuesday, October 10th.</h1>
−
  <ul>
+
−
    <li>Digestions of Bba_1, Bba_2 and Bba_3, 08:00.</li>
+
−
    <ul>
+
−
      <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
    <li>Digestion of the modified phage, 09:00.</li>
+
−
    <ul>
+
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
    <li>Electrophoresis, 12:00.</li>
+
−
    <ul>
+
−
      <li>Eduardo Ramirez (2).</li>
+
−
    </ul>
+
−
    <li>Ligation of the phages back to circle form, 13:00.</li>
+
−
    <ul>
+
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
  </ul>
+
 
</div>
 
</div>
  
 
<div id="date-10-11">
 
<div id="date-10-11">
−
   <h1>Saturday, October 11th.</h1>
+
   <h1>Wednesday, October 11th.</h1>
−
  <ul>
+
−
    <li>Inoculations for constructs stocks, 20:00.</li>
+
−
    <ul>
+
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
  </ul>
+
 
</div>
 
</div>
  
 
<div id="date-10-12">
 
<div id="date-10-12">
−
   <h1>Sunday, October 12th.</h1>
+
   <h1>Thursday, October 12th.</h1>
−
  <ul>
+
       <li>Tatenda Tela</li>
−
    <li>Pictures taken for the wiki, 13:00.</li>
+
<ul>I transformed HCG in PS1C3 into E.coli today</ul>
−
    <ul>
+
<li> Cara Jones</li>
−
       <li>Everyone.</li>
+
<ul>PCR Amplification, Restriction Digest: <a href="https://static.igem.org/mediawiki/2017/e/e0/T--Georgia_State--10_12_17_cj.pdf"> See attached procedure for instructions.</a>  
−
    </ul>
+
−
    <li>PCR, 16:00.</li>
+
−
    <ul>
+
−
      <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
    <li>Stocks, 18:00.</li>
+
−
    <ul>
+
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
    <li>Electrophoresis, 20:00.</li>
+
−
    <ul>
+
−
      <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
  </ul>
+
 
</div>
 
</div>
 +
  
 
<div id="date-10-13">
 
<div id="date-10-13">
−
   <h1>Monday, October 13th.</h1>
+
   <h1>Friday, October 13th.</h1>
−
  <ul>
+
       <li>Tatenda Tela</li>
−
    <li>PCR for Red Lambda confirmation, 13:00.</li>
+
<ul>I performed PCR and did overnight cultures of 10 colonies</ul>
−
    <ul>
+
−
       <li>Eduardo Cepeda.</li>
+
−
    </ul>
+
−
    <li>Electrophoresis, 15:00.</li>
+
−
    <ul>
+
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
    <li>Digestions, 16:00.</li>
+
−
    <ul>
+
−
      <li>Mercedes.</li>
+
−
    </ul>
+
−
    <li>TAE buffer preparation, 20:00.</li>
+
−
    <ul>
+
−
      <li>Newbies: Alejandro, Danny.</li>
+
−
    </ul>
+
−
    <li>Western Blott, 21:00.</li>
+
−
    <ul>
+
−
      <li>Minerva, Claudia.</li>
+
−
    </ul>
+
−
  </ul>
+
 
</div>  
 
</div>  
  
 
<div id="date-10-14">
 
<div id="date-10-14">
−
   <h1>Tuesday, October 14th.</h1>
+
   <h1>Saturday, October 14th.</h1>
−
  <ul>
+
      <li>Eudoxie Bataba</li>
−
    <li>Work on wiki</li>
+
<ul><b>Objective:</b>PCR diagnostic gel, Miniprep and Restriction DIgest</ul>
−
    <ul>
+
<ul><b>Materials:</b>QIAGEN miniprep kit, 50% glycerol</ul>
−
      <li>Sebasti&aacute;n Valdivieso, Eduardo Zardain, Eduardo Ram&iacute;rez</li>
+
<ul><b>Equipment:</b>Centrifuge, Nanophotometer, Thermocycler</ul>
−
    </ul>
+
<ul><b>Procedure:</b><a href="https://2017.igem.org/File:T--Georgia_State--HCG_PSB.jpg">See attached procedure.</a></ul>  
−
  </ul>
+
<ul><b>Result:</b><a href="https://2017.igem.org/File:T--Georgia_State--_PCR_Oct_14.jpg">See attached PCR results.</a></ul>
 +
<ul><b>Result:</b><a href="https://2017.igem.org/File:T--Georgia_State--_Miniprep_Oct_14.jpg">See attached miniprep results.</a></ul>
 +
<ul><b>Result:</b><a href="https://2017.igem.org/File:T--Georgia_State--_RD_Oct_14.jpg">See attached Restriction Digest results.</a></ul>  
 
</div>
 
</div>
 +
  
 
<div id="date-10-15">
 
<div id="date-10-15">
−
   <h1>Wednesday, October 15th.</h1>
+
   <h1>Sunday, October 15th.</h1>
−
  <ul>
+
<li>Tatenda Tela</li>
−
    <li>Work on wiki</li>
+
<ul>Restriction digests</ul>
−
    <ul>
+
<ul>Ligation</ul>
−
      <li>Sebasti&aacute;n Valdivieso, Eduardo Zardain, Eduardo Ram&iacute;rez</li>
+
−
    </ul>
+
−
  </ul>
+
 
</div>
 
</div>
  
 
<div id="date-10-16">
 
<div id="date-10-16">
−
  <h1>Thursday, October 16th.</h1>
+
<h1>Monday,October 16th.</h1>
−
  <ul>
+
<li>Eudoxie Bataba</li>
−
    <li>Work on wiki</li>
+
<ul> Transformations</ul>
−
    <ul>
+
<li> Cara Jones</li>
−
      <li>Sebasti&aacute;n Valdivieso, Eduardo Zardain, Eduardo Ram&iacute;rez</li>
+
<ul>PCR amplification of mamba in pGEX, Digest of amplified mamba: <a href="https://static.igem.org/mediawiki/2017/0/08/T--Georgia_State--10_16_17_cj.pdf"> See attached procedure for instructions.</a>  
−
    </ul>
+
−
  </ul>
+
 
</div>
 
</div>
−
 
 
<div id="date-10-17">
 
<div id="date-10-17">
−
   <h1>Friday, October 17th.</h1>
+
   <h1>Tuesday, October 17th.</h1>
−
   <ul>
+
<li>Tatenda Tela</li>
−
    <li>Work on wiki, its wiki-freeze day!!!</li>
+
<ul>Ligation</ul>
−
    <ul>
+
</div>
−
      <li>Sebasti&aacute;n Valdivieso, Eduardo Zardain, Eduardo Ram&iacute;rez</li>
+
<div id="date-10-18">
−
    </ul>
+
<h1>Wednesday,October 18th.</h1>
−
   </ul>
+
<li>Tatenda Tela and Cara Jones</li>
 +
<ul>Digested miniprepped DNA from glycerol stocks</ul>
 +
<ul>Gel extraction of the backbones</ul>
 +
<br>
 +
<li> Cara Jones</li>
 +
<ul>PCR of amplified mamba+prefix and suffix: <a href="https://static.igem.org/mediawiki/2017/c/c8/T--Georgia_State--10_18_17_cj.pdf"> See attached procedure for instructions.</a>
 +
</div>
 +
<div id="date-10-19">
 +
<h1> Thursday, October 19th </h1>
 +
<li>Tatenda Tela</li>
 +
<ul>Double digests</ul>
 +
<ul>Ligations</ul>
 +
<ul>Transformations</ul>
 +
<br>
 +
<li> Cara Jones</li>
 +
<ul>PCR cleanup of mamba: <a href="https://static.igem.org/mediawiki/2017/6/68/T--Georgia_State--10_19_17_cj.pdf"> See attached procedure for instructions.</a> 
 +
</div>
 +
<div id="date-10-20">
 +
<h1> Friday, October 20th </h1>
 +
<li> Tatenda Tela</li>
 +
<ul>Colony pcr</ul>
 +
<ul><table><tr><td><img src="https://static.igem.org/mediawiki/2017/c/ca/T--Georgia_State--_plates_of_hcg_in_psb_with_colonies_picked.jpeg" alt=""></td><tr><td><img src="https://static.igem.org/mediawiki/2017/c/cb/T--Georgia_State--mamba_in_pSB1C3_plates_with_colonies_picked.jpeg" alt=""></td></tr></table></ul>
 +
</div>
 +
<div id="date-10-21">
 +
<h1> Saturday, October 21st </h1>
 +
<li>Tatenda Tela and Aditya Natu</li>
 +
<ul>Miniprep colonies</ul>
 +
</div>
 +
<div id="date-10-22">
 +
<h1>Sunday, October 22nd.</h1>
 +
</div>
 +
<div id="date-10-23">
 +
   <h1>Monday, October 23rd.</h1>
 +
<li>Tatenda Tela and Aditya Natu </li>
 +
<ul>Colony PCR</ul>  
 +
<ul> Diagnostic restriction digest of colonies </ul>
 +
<ul> Gel extraction and clean-up</ul>
 +
</div>
 +
<div id="date-10-24">
 +
  <h1>Tuesday, October 24th.</h1>
 +
<li>Tatenda Tela </li>
 +
<ul> Digested three different plasmids for ligation with HCG. All were PSB1C3 with different inserts.</ul>
 +
<ul> Gel extraction and clean-up</ul>
 +
</div>
 +
<div id="date-10-25">
 +
  <h1>Wednesday, October 25th.</h1>
 +
<li> Tatenda Tela>
 +
<ul> Minipreps of colonies from the plates from 10/19</ul>
 +
<ul> Restriction digests</ul>
 +
</div>
 +
<div id="date-10-26">
 +
  <h1>Thursday, October 26th.</h1>
 +
<li> Tatenda Tela</li>
 +
<ul> Diagnostic gel </ul>
 +
<ul> Minipreps </ul>
 +
</div>
 +
<div id="date-10-27">
 +
  <h1>Friday, October 27th.</h1>
 +
<strong> DNA Submission Deadline!!</strong>
 +
</div>
 +
<div id="date-10-28">
 +
  <h1>Saturday, October 28th.</h1>
 +
</div>
 +
<div id="date-10-29">
 +
  <h1>Sunday, October 29th.</h1>
 +
</div>
 +
<div id="date-10-30">
 +
  <h1>Monday, October 30th.</h1>
 +
<li> Everyone </li>
 +
<ul><strong>Freak out about our wiki.</strong></ul>
 +
<br>
 +
<li> Cara Jones</li>
 +
<ul>Mamba in pGEX concentration, mamba in pGEX column chromatography, HRV cleavage of mamba-gst fusion, SDS PAGE: <a href="https://static.igem.org/mediawiki/2017/5/5c/T--Georgia_State--10_30_31_17_cj.pdf"> See attached procedure for instructions.</a>
 +
</div>
 +
 
 +
<div id="date-10-31">
 +
  <h1>Tuesday, October 31st.</h1>
 +
<li> Everyone </li>
 +
<ul><strong>Freak out about our wiki.</strong></ul>
 +
</div>
 +
 
 +
<div id="date-11-01">
 +
   <h1>Wednesday, November 1st.</h1>
 +
<strong> WIKI FREEZE!!!!!!</strong>
 +
<li> Everyone</li>
 +
<ul><strong>Freak out about our wiki.</strong></ul>
 
</div>
 
</div>
  

Latest revision as of 03:58, 2 November 2017

<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" "http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd"> f Team:Tec-Monterrey/ITESM14 notebook data.html - 2014.igem.org

Monday, May 1st.

Sunday, June 1st.

Monday, June 2nd.

Tuesday, June 3rd.

Wednesday, June 5th.

Thursday, June 5th.

Friday, June 6th.

Saturday, June 7th.

Sunday, June 8th.

Monday, June 9th.

Tuesday, June 10th.

Wednesday, June 11th.

Thursday, June 12th.

Friday, June 13th.

Saturday, June 14th.

Sunday, June 15th.

Monday, June 16th.

Tuesday, June 17th.

Wednesday, June 18th.

Thursday, June 19th.

Friday, June 20th.

Saturday, June 21st.

Sunday, June 22nd.

Monday, June 23rd.

Tuesday, June 24th.

Wednesday, June 25nd.

Thursday, June 26th.

Friday, June 27th.

  • Cara Jones

    • Saturday, June 28nd.

    Sunday, June 29nd.

    Monday, June 30nd.

    Tuesday, July 1st.

    Wednesday, July 2nd.

    Thursday, July 3rd.

    Friday, July 4th.

    Wednesday, July 5th.

    Thursday, July 6th.

  • Cara Jones

    • Friday, July 7th.

  • Tori Radcliff
      • Objective:To determine if Alkaline Phosphotase will bind to nitrocellulose paper and produce a signal when mixed with BCIP.
      Materials:Cut small strips of nitrocellulose paper (Nitran), Promega Alkaline Phosphatase (Conc: 1ng/ul), Alkaline 10X buffer diluted, BCIP
      Equipment:UV-Crosslinker
      Results: The Ap spread outward but stayed confined meaning the crosslinking worked. Adding the BCIP solution to the tube with the paper produced a noticeable discoloration. Adding the enzyme to the BCIP only produced a small discoloration. The binding of the alkaline phosphatase and BCIP to the paper was successful. The reaction worked only in the intended areas meaning that if the AP is bound to the factor c it may produce a signal only if it is cut, the AP nor BCIP disperses in the solution after being bound to the paper.
  • Cara Jones

    • Saturday, July 8th.

    Sunday, July 9th.

    Monday, July 10th.

  • Cara Jones

    • Tuesday, July 11th.

  • Cara Jones

    • Wednesday, July 12th.

    Thursday, July 13th.

  • Cara Jones

    • Friday, July 14th.

    Saturday, July 15th.

    Sunday, July 16th.

    Monday, July 17th.

  • Cara Jones

    • Tuesday, July 18th.

    Wednesday, July 19th.

    Thursday, July 20th.

    Friday, July 21st.

    Saturday, July 22nd.

    Sunday, July 23rd.

    Monday, July 24th.

    Tuesday, July 25th.

    Wednesday, July 26th.

    Thursday, July 27th.

  • Cara Jones

    • Friday, July 28th.

    Saturday, July 29th.

    Sunday, July 30th.

    Monday, July 31st.

    Tuesday,August 1st.

  • Tori Radcliff
      • Objective:Before the fusion step, the hCG needs to be placed into the PSB1C3 plasmid, so that the hCG can be submitted. In order to add the hCG to add to the plasmid restriction sites need to be added this was done by adding the prefix and suffix through PCR.
      Materials:Qiagen Master mix (2x), Prefix Primer, Suffix Primer, hCG DNA (diluted to 10ng/ul), and Nuclease free H20
      Equipment:Thermocycler
      Results:
      Simulated gel using SnapGene1% Agarose gel, experimental results

    Wednesday, August 2nd.

    Thursday, August 3rd.

  • Tori Radcliff
      • Objective:To double digest both the PSB1C3 backbone and our hCG PCR product
      Materials:DNA of ECFP-PCB1C3 (102 ng/ul) and hCG PCR product, Restriction enzymes: NotI, EcoRI, XbaI, PstI,SpeI, Green Fastdigest Buffer (10x), and Nuclease free H20
      Equipment:Thermocycler
      Results:
      Simulated gel using SnapGeneThe Wells Contain the following:
    • 1. 1Kb New England Biolabs Ladder
    • 2. hCG uncut
    • 3. hCG cut with EcoR1 and Pst1
    • 4. hCG cut with Xba1 and Spe1
    • 5. ECFP cut with EcoR1 and Pst1
    • 6. ECFP cut with Xba1 and Spe1

    Friday August 4th.

  • Cara Jones
      • Objective: Clean used GST resin so that it may be used in future protein purifications.
      Materials: GST Resin, 70% Ethanol, GST Elution Buffer
      Equipment: Centrifuge,Vortexer, Micropipette,Eppendorf tubes/Falcon tubes, Spectrophotometer, and Cuvettes

    Saturday, August 5th.

  • Tori Radcliff
      • Objective:To repeat the double digest of PSCB1C3 and hCG PCR
      Materials:DNA of Lt10-PCB1C3 (80 ng/ul) and hCG PCR product, Restriction enzymes: NotI, EcoRI, XbaI, PstI,SpeI, Green Fastdigest Buffer (10x), and Nuclease free H20
      Equipment:Thermocycler
      Results:
      Since the diagnostic gel was a success the PSB1C backbone was extracted from the Lt 10 plasmid using an E-gel.

    Sunday, August 6th.

    Monday, August 7th.

  • Cara Jones
      • Objective:Culture of transformed E.coli with mamba in PGEX
      I picked 10 colonies from plates and did overnight cultures in LB

    Tuesday, August 8th.

  • Eudoxie Bataba
      • Objective:I will get the mamba in PGEX plasmid from transformed E.coli
      Materials:NEB miniprep kit, 50% glycerol
      Equipment:Centrifuge and Nanophotometer
      Results:
      Sample Concentration (ng/uL)
      1 118
      2 123
      3 85
      4 133
      5 90
      6 75
      7 72
      8 100
      9 65
      10 140

  • Cara Jones

    • Wednesday, August 9th.

  • Tori Radcliff
      • Ligation of hCG to PSB1C3

    Thursday, August 10th.

  • Tori Radcliff
      • Transformation of hCG-PSB1C3 ligation and overnight culture

    Friday, August 11th.

  • Tori Radcliff
      • Colony PCR of hCG-PSB1C3 colonies

    Saturday, August 12th.

    Sunday, August 13th.

  • Tori Radcliff
      • Overnight culture of hCG-PSB1C3 colonies

    Monday, August 14th.

  • Tori Radcliff
      • Miniprep of hCG-PSB1C3 cultures
      Diagnostic restriction digest
      PCR of factor-C gBlocks
        Objective: Factor C was divided into 4 G-blocks and ordered from IDT. A PCR was performed to add overlaps to each block so that subsequent overlap extension PCRs could be performed.
        Results:
        A simulated gel was created and then the actual sampled of each were run on an e-gel. 5ul of product with 1ul of 6X dye and 14 ul of DI water was added to each well. The wells include the following from left to right: Gene Ruler 1kb DNA ladder, PCR of g-block 1, PCR of g-block 2, PCR of g-block 3, PCR of g-block 4.
    • Cara Jones

    Tuesday, August 15th.

  • Tori Radcliff
      • Overlap Extension PCR
      Objective:To combine the four PCR-ed g-blocks of factor-C.
      Results:
      Simulated gelWells contain the following: Gene Ruler 1kb plus MW ladder, Reaction of blocks 1 and 2, Negative control (blocks 1 and 2), Reaction of blocks 3 and 4, Negative control (blocks 3 and 4), Reaction of 1 and 2 with 3 and 4 products, negative control (reactions 1 and 2 with 3 and 3 and 4), and Gene Ruler 1kb plus MW ladder.

    Wednesday, August 16th.

    Thursday, August 17th.

  • Tori Radcliff
      • Objective:Prepare hCG for ligation into pGEX through PCR
      Materials:hCG DNA, primers forward and reverse and master mix
      Equipment:Thermocycler
      Procedure:Two 50 ul PCR reactions were performed. 25ul of qiagen mastermix, 2ul of block 5 DNA, 2.5ul of forward primer, 2.5 ul of reverse primer and 18 ul of nuclease-free water was added to tube 1 and tube 2.
  • Tatenda Tela
      • Overnight cultures
      Colony PCR of Cara's transformants

    Friday, August 18th.

    Saturday, August 19th.

    Sunday, August 20th.

    Monday, August 21st.

  • Tatenda Tela
      • Miniprep
      Ligations

    Tuesday, August 22nd.

    Wednesday, August 23rd.

  • Cara Jones

    • Thursday, August 24th.

  • Cara Jones

    • Friday, August 25th.

  • Tori Radcliff
      • Objective:To repeat the double digest of PSCB1C3 and hCG PCR
      Materials:DNA of Lt10-PCB1C3 (80 ng/ul) and hCG PCR product, Restriction enzymes: NotI, EcoRI, XbaI, PstI,SpeI, Green Fastdigest Buffer (10x), and Nuclease free H20
      Equipment:Thermocycler
  • Cara Jones

    • Saturday, August 26th.

    Sunday, August 27th.

    Monday, August 28th.

  • Cara Jones

    • Tuesday, August 29th.

  • Tori Radcliff
      • Ligation of hCG and PSB1C3
        Objective:To ligate hCG and PSB1C3
        Materials: digested vector cleaned by Cara Jones, T4 ligase and buffer, digested hCG
        Procedure: Using a ligation calculator the concentration needed was 31.87 ng/ul in a 20ul total ligation. 1ul of PGEX at 105 ng/ul and 6 ul of hCG, 2 ul of T4 buffer, 1 ul of T4 ligase and 10 ul of nuclease-free water. A negative control was used which included digested mamba in pgex with no ligation buffer. The ligation reaction sat at room temperature for 10 minutes.
      Transformation
        Objective:To transform the ligation of hCG and PSB1C3
        Materials:C2987I NEB competent cells, ligation reaction
        Equipment:shaker and incubator
        Results:There was no growth on the ligation plate nor negative control but there was growth on the positive control. Since there was no growth on the ligation plates that would mean the cells have no resistance to ampicillin and the ligation was inefficient due to the fact the hCG was not purified to eliminate contaminants ike EDTA and salts.

    Wednesday, August 30th.

  • Cara Jones

    • Thursday, August 31st.

    Friday, September 1st.

    Saturday, September 2nd.

    Sunday, September 3rd

    Monday, September 4th

    Tuesday, September 5th

    Wednesday, September 6

    Thursday, September 7

    Friday, September 8th.

  • Tatenda Tela
      • hCG for pGEX pcr
      Results:
      lanecontents
      1Generuler 1kb plus ladder
      2hCG insert w/ hCG block 5 fwd&rev primers
      3hCG insert w/ hCG block 5 fwd&rev primers
      4hCG insert w/ hCG block 5 fwd&rev primers
      5hCG insert w/ hCG block 5 fwd&rev primers
      6hCG insert w/ hCG block 5 fwd&rev primers
      7hCG insert w/ hCG block 5 fwd&rev primers
      8hCG insert w/ hCG for pGEX fwd&rev primers
      9hCG insert w/ hCG for pGEX fwd&rev primers
      10hCG insert w/ hCG for pGEX fwd&rev primers
      11hCG insert w/ hCG for pGEX fwd&rev primers
      12hCG insert w/ hCG for pGEX fwd&rev primers
      13hCG insert w/ hCG for pGEX fwd&rev primers
      14Generuler 1kb plus ladder

    Saturday, September 9

    Sunday, September 10

    Monday, September 11

    Tuesday, September 12

    Wednesday, September 13

  • Cara Jones
      • Overnight Cultures
  • Tatenda Tela
      • Restriction digest and heat inactivation of samples from September 8

    Thursday, September 14th.

  • Tatenda Tela
      • Miniprep of overnight cultures prepped by Cara the day before
      Results:
      Sample Concentration (ng/uL) A260/A280
      1 252.0
      2 7.52.0
      3 9.8 1.7
      4 8.5 1.8
      5 9.5 1.7

  • Cara Jones

    • Friday, September 15

    Saturday, September 16th.

  • Victoria Radcliffe, Aditya Natu
      • Midiprep of mamba in pGEX to get pGEx backbone
    sampleconcentration (ng/uL)
    1200
    2275

    Sunday, September 17

    Monday, September 18

    Tuesday, September 19

  • Tatenda Tela
      • Digest of Midiprep of mamba in pGEX done by Victoria and Aditya using EcoRI and NotI enzymes
      Results:
      lanecontents
      1Generuler 1kb plus ladder
      2Mamba in pGEX
      3Mamba in pGEX
      4Mamba in pGEX cut with EcoRI
      5Mamba in pGEX cut with EcoRI
      6empty
      7Mamba in pGEX cut with EcoRI and NotI
      8empty
      9Mamba in pGEX cut with EcoRI and NotI
      10empty
      11Mamba in pGEX cut with EcoRI and PstI
      12Mamba in pGEX cut with EcoRI and PstI
      13empty
      14empty
  • Tatenda Tela
      • Gel extraction of pGEX backbone (lanes 7 and 9)
      Ligation of digested pGEX backbone and digested hCG from September 13
      Transforamtion using 5 alpha competent cells

    Wednesday, September 20

    Thursday, September 21

  • Tatenda Tela
      • Another digest mamba in pGEX and then a ligation with the hCG insert followed by a transformation into 5 alpha competent cells

    Friday, September 22

  • Julia Ngo
    • PCR of hCG using hCG for pGEX fwd and rev primers

    • Saturday, September 23

    Sunday, September 24

    Monday, September 25

  • Julia Ngo
      • Ran gel of hCG PCR from September 22

    Tuesday, September 26

    Wednesday, September 27

    Thursday, September 28

    Friday, September 29

  • Julia Ngo
      • Designing Primers for Tac-GST-Mamba in pSB1C3

    Saturday, September 30

    Sunday, October 1st.

    Monday, October 2nd.

  • Cara Jones

    • Tuesday, October 3rd.

  • Julia Ngo
      • PCR of Tac promoter

    Wednesday, October 4th.

  • Cara Jones

    • Thursday, October 5th.

    Friday, October 6th.

    Saturday, October 7th.

    Sunday, October 8th.

    Monday, October 9th.

  • Cara Jones

    • Tuesday, October 10th.

    Wednesday, October 11th.

    Thursday, October 12th.

  • Tatenda Tela
      • I transformed HCG in PS1C3 into E.coli today
  • Cara Jones

    • Friday, October 13th.

  • Tatenda Tela
      • I performed PCR and did overnight cultures of 10 colonies

    Saturday, October 14th.

  • Eudoxie Bataba
      • Objective:PCR diagnostic gel, Miniprep and Restriction DIgest
      Materials:QIAGEN miniprep kit, 50% glycerol
      Equipment:Centrifuge, Nanophotometer, Thermocycler

    Sunday, October 15th.

  • Tatenda Tela
      • Restriction digests
      Ligation

    Monday,October 16th.

  • Eudoxie Bataba
      • Transformations
  • Cara Jones

    • Tuesday, October 17th.

  • Tatenda Tela
      • Ligation

    Wednesday,October 18th.

  • Tatenda Tela and Cara Jones
      • Digested miniprepped DNA from glycerol stocks
      Gel extraction of the backbones

  • Cara Jones

    • Thursday, October 19th

  • Tatenda Tela
      • Double digests
      Ligations
      Transformations

  • Cara Jones

    • Friday, October 20th

  • Tatenda Tela
      • Colony pcr

    Saturday, October 21st

  • Tatenda Tela and Aditya Natu
      • Miniprep colonies

    Sunday, October 22nd.

    Monday, October 23rd.

  • Tatenda Tela and Aditya Natu
      • Colony PCR
      Diagnostic restriction digest of colonies
      Gel extraction and clean-up

    Tuesday, October 24th.

  • Tatenda Tela
      • Digested three different plasmids for ligation with HCG. All were PSB1C3 with different inserts.
      Gel extraction and clean-up

    Wednesday, October 25th.

  • Tatenda Tela>
      Minipreps of colonies from the plates from 10/19
      Restriction digests

    • Thursday, October 26th.

  • Tatenda Tela
      • Diagnostic gel
      Minipreps

    Friday, October 27th.

    DNA Submission Deadline!!

    Saturday, October 28th.

    Sunday, October 29th.

    Monday, October 30th.

  • Everyone
      • Freak out about our wiki.

  • Cara Jones

    • Tuesday, October 31st.

  • Everyone
      • Freak out about our wiki.

    Wednesday, November 1st.

    WIKI FREEZE!!!!!!
  • Everyone
      • Freak out about our wiki.

    Retrieved from "https://2014.igem.org/Team:Tec-Monterrey/ITESM14_notebook_data.html"