Difference between revisions of "Team:Northwestern/08 26"

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     <h1>Friday, August 26<sup>th</sup></h3>
 
     <h1>Friday, August 26<sup>th</sup></h3>
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    <h2>Agenda:</h2>
+
<h2>Tasks:</h2>
 +
    <div class="row">
 +
      <div class="col-sm-2">
 +
        <p>Michelle</p>
 +
      </div>
 +
      <div class="col-sm-10">
 +
        <ul>
 +
          <li>Gel extracted gRNA and signal sequences</li>
 +
          <ul><li>Used IBI protocol (kit from Leonard Lab)</li>
 +
          <li>Nanodrop results: all under 10 ng/uL, 260/280 above 1.9, 260/230 around 0.5 — discarded </li></ul>
 +
          <li>Reran signal sequence PCR (to add homology)</li>
 +
          <ul><li>1 uL template</li>
 +
          <li>1 uL fwd primer 10uM</li>
 +
          <li>1 uL rev primer 10uM</li>
 +
          <li>1 uL DMSO</li>
 +
          <li>21 uL nfH<sub>2</sub>O</li>
 +
          <li>25 uL OneTaq Master Mix </li></ul>
 +
          <li>Ran 2 gels on the signal sequence PCR products
 +
            <div class="row">
 +
              <div class="col-xs-8"><img src="https://static.igem.org/mediawiki/2016/7/79/T--Northwestern--08_26_1.png" width="1134" height="230" alt=""/></div>
 +
            </div>
 +
            <div class="row">
 +
              <div class="col-xs-6"><img src="https://static.igem.org/mediawiki/2016/e/e7/T--Northwestern--08_26_2.jpg" width="1134" height="230" alt=""/></div>
 +
              <div class="col-xs-6"><img src="https://static.igem.org/mediawiki/2016/0/03/T--Northwestern--08_26_3.jpg" width="1134" height="230" alt=""/></div>
 +
            </div>
 +
          </li>
 +
        </ul>
 +
      </div>
 +
    </div>
 +
    <div class="row">
 +
      <div class="col-sm-2">
 +
        <p>Tasfia</p>
 +
      </div>
 +
      <div class="col-sm-10">
 +
        <ul>
 +
          <li>DpnI digested Cas9-Lrz-SS PCR product</li>
 +
          <li>Made more 10X TAE</li>
 +
          <li>Registered all the ChemEs and Jordan for iGEM Jamboree</li>
 +
          <li>DpnI digested Tet-Lrz-GFP for two hours and ran a gel screen on it (with 1 uL of our remaining SybrGreen)
 +
            <div class="row">
 +
              <div class="col-xs-6"><img src="https://static.igem.org/mediawiki/2016/8/88/T--Northwestern--08_26_4.png" width="242" height="418" alt=""/></div>
 +
            </div>
 +
          </li>
 +
          <ul>
 +
            <li>Only primer dimers result from the Tet-Lrz-GFP PCR </li>
 +
          </ul>
 +
        </ul>
 +
      </div>
 +
    </div>
 +
    <div class="row">
 +
      <div class="col-sm-2">
 +
        <p>Tyler</p>
 +
      </div>
 +
      <div class="col-sm-10">
 +
        <ul>
 +
          <li>Gibson Assembly of gRNA-mRFP</li>
 +
          <ul>
 +
            <li>gRNA-mRFP Assembly</li>
 +
            <ul>
 +
              <li>0.31 µL insert</li>
 +
              <li>0.4 µL backbone</li>
 +
              <li>4.29 µL water</li>
 +
              <li>5 µL Gibson mix</li>
 +
            </ul>
 +
            <li>Negative Control</li>
 +
            <ul>
 +
              <li>0.4 µL backbone</li>
 +
              <li>4.6 µL water</li>
 +
              <li>5 µL Gibson Mix</li>
 +
            </ul>
 +
            <li>Quantities transformed (heat shocked 30s, 200µL SOC)</li>
 +
            <ul>
 +
              <li>3 µL gRNA-mRFP assembly product </li>
 +
              <li>2 µL negative control</li>
 +
              <li>2µL positive Gibson kit control</li>
 +
              <li>1µL positive transformation control</li>
 +
            </ul>
 +
          </ul>
 +
        </ul>
 +
      </div>
 +
    </div>
 +
  </article>
  
 
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Revision as of 00:19, 16 October 2016

Notebook

Friday, August 26th

Tasks:

Michelle

  • Gel extracted gRNA and signal sequences
    • Used IBI protocol (kit from Leonard Lab)
    • Nanodrop results: all under 10 ng/uL, 260/280 above 1.9, 260/230 around 0.5 — discarded
  • Reran signal sequence PCR (to add homology)
    • 1 uL template
    • 1 uL fwd primer 10uM
    • 1 uL rev primer 10uM
    • 1 uL DMSO
    • 21 uL nfH2O
    • 25 uL OneTaq Master Mix
  • Ran 2 gels on the signal sequence PCR products

Tasfia

  • DpnI digested Cas9-Lrz-SS PCR product
  • Made more 10X TAE
  • Registered all the ChemEs and Jordan for iGEM Jamboree
  • DpnI digested Tet-Lrz-GFP for two hours and ran a gel screen on it (with 1 uL of our remaining SybrGreen)
    • Only primer dimers result from the Tet-Lrz-GFP PCR

Tyler

  • Gibson Assembly of gRNA-mRFP
    • gRNA-mRFP Assembly
      • 0.31 µL insert
      • 0.4 µL backbone
      • 4.29 µL water
      • 5 µL Gibson mix
    • Negative Control
      • 0.4 µL backbone
      • 4.6 µL water
      • 5 µL Gibson Mix
    • Quantities transformed (heat shocked 30s, 200µL SOC)
      • 3 µL gRNA-mRFP assembly product
      • 2 µL negative control
      • 2µL positive Gibson kit control
      • 1µL positive transformation control