Difference between revisions of "Team:Northwestern/08 29"

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     <h1>Monday, August 29<sup>th</sup></h3>
 
     <h1>Monday, August 29<sup>th</sup></h3>
 
     <h2>Agenda:</h2>
 
     <h2>Agenda:</h2>
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<h2>Tasks:</h2>
 +
    <div class="row">
 +
      <div class="col-sm-2">
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        <p>Tasfia</p>
 +
      </div>
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      <div class="col-sm-10">
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        <ul>
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          <li>Made 5 mL overnight cultures of "TetR-Cas9," "Assembled Cas9 from Cam Culture," GFP from iGEM kit</li>
 +
          <ul>
 +
            <li>Three different colonies</li>
 +
            <li>Used 1000X Cam</li>
 +
            <li>Working concentration was 35 ug/mL</li>
 +
            <li>Added 5.15 uL to each culture tube </li>
 +
          </ul>
 +
          <li>Made 5 mL overnight culture of gRNA Gibson product</li>
 +
          <ul>
 +
            <li>One colony, duplicate overnight cultures</li>
 +
            <li>Used 1000X Tet (10 mg/mL)</li>
 +
            <li>Working concentration was 10 ug/mL</li>
 +
            <li>Added 5 uL to each culture tube</li>
 +
          </ul>
 +
          <li>Gel extracted signal sequences (with homology added)
 +
            <div class="row">
 +
              <div class="col-xs-8 col-sm-12"><img src="https://static.igem.org/mediawiki/2016/5/50/T--Northwestern--08_28_1.png" width="1210" height="440" alt=""/></div>
 +
            </div>
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          </li>
 +
          <li>Used team-modified protocol:</li>
 +
          <ul>
 +
            <li>Sat product for ~4 minutes between washes</li>
 +
            <li>Evaporated EtOH for ~12 minutes</li>
 +
            <li>Eluted in nuclease-free water after standing for ~15 minutes</li>
 +
          </ul>
 +
          <li>Going forward: we can attempt a Cas9-SS Gibson after figuring out what’s wrong with the gRNA Gibson assembly</li>
 +
        </ul>
 +
      </div>
 +
    </div>
 +
    <div class="row">
 +
      <div class="col-sm-2">
 +
        <p>Tyler</p>
 +
      </div>
 +
      <div class="col-sm-10">
 +
        <ul>
 +
          <li>Retransformed gRNA Gibson product from 08.26.16 </li>
 +
          <ul>
 +
            <li>3 µL of gRNA Gibson assembly product</li>
 +
            <li>2 µL positive Gibson kit control</li>
 +
            <li>2 µL (-) control 1 µL transformation control</li>
 +
          </ul>
 +
          <li>Retransformed 1 µL GFP from iGEM kit</li>
 +
        </ul>
 +
      </div>
 +
    </div>
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  </article>
 
<footer id="nav">
 
<footer id="nav">
 
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     </footer>
 
     </footer>
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<script type="text/javascript" src="https://2016.igem.org/Team:Northwestern/libraries/jquery?action=raw&ctype=text/javascript"></script>
 
<script type="text/javascript" src="https://2016.igem.org/Team:Northwestern/libraries/jquery?action=raw&ctype=text/javascript"></script>
 
         <script type="text/javascript" src="https://2016.igem.org/Team:Northwestern/libraries/bootstrap?action=raw&ctype=text/javascript"></script>
 
         <script type="text/javascript" src="https://2016.igem.org/Team:Northwestern/libraries/bootstrap?action=raw&ctype=text/javascript"></script>

Revision as of 00:27, 16 October 2016

Notebook

Monday, August 29th

Agenda:

Tasks:

Tasfia

  • Made 5 mL overnight cultures of "TetR-Cas9," "Assembled Cas9 from Cam Culture," GFP from iGEM kit
    • Three different colonies
    • Used 1000X Cam
    • Working concentration was 35 ug/mL
    • Added 5.15 uL to each culture tube
  • Made 5 mL overnight culture of gRNA Gibson product
    • One colony, duplicate overnight cultures
    • Used 1000X Tet (10 mg/mL)
    • Working concentration was 10 ug/mL
    • Added 5 uL to each culture tube
  • Gel extracted signal sequences (with homology added)
  • Used team-modified protocol:
    • Sat product for ~4 minutes between washes
    • Evaporated EtOH for ~12 minutes
    • Eluted in nuclease-free water after standing for ~15 minutes
  • Going forward: we can attempt a Cas9-SS Gibson after figuring out what’s wrong with the gRNA Gibson assembly

Tyler

  • Retransformed gRNA Gibson product from 08.26.16
    • 3 µL of gRNA Gibson assembly product
    • 2 µL positive Gibson kit control
    • 2 µL (-) control 1 µL transformation control
  • Retransformed 1 µL GFP from iGEM kit