Difference between revisions of "Team:Northwestern/08 31"

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     <h1>Wednesday, August 31<sup>st</sup></h3>
 
     <h1>Wednesday, August 31<sup>st</sup></h3>
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    <h2>Agenda:</h2>
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  <h2>Tasks:</h2>
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    <div class="row">
 +
      <div class="col-sm-2">
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        <p>Michelle</p>
 +
      </div>
 +
      <div class="col-sm-10">
 +
        <ul>
 +
          <li>Miniprepped gRNA overnight colony</li>
 +
          <ul>
 +
            <li>216.0 ng/uL, 260/280: 1.87, 260/230: 2.20</li>
 +
          </ul>
 +
          <li>Made nfH2O aliquots</li>
 +
          <li>Autoclaved things</li>
 +
          <li>Grew up TetR-Cas9 for western blot</li>
 +
          <ul>
 +
            <li>50 mL culture</li>
 +
            <li>Starting OD<sub>600</sub>: 0.035 A</li>
 +
            <li>1 hour OD<sub>600</sub>: 0.048 A</li>
 +
            <li>2 hour OD<sub>600</sub>: 0.157 A</li>
 +
            <li>2.5 hour OD<sub>600</sub>: 0.249 A</li>
 +
            <li>3 hour OD<sub>600</sub>: 0.464 A</li>
 +
            <li>3.25 hr OD<sub>600</sub>: 0.600 A</li>
 +
          </ul>
 +
        </ul>
 +
      </div>
 +
    </div>
 +
    <div class="row">
 +
      <div class="col-sm-2">
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        <p>Tasfia</p>
 +
      </div>
 +
      <div class="col-sm-10">
 +
        <ul>
 +
          <li>Gel extracted PCR products (stored in -20&#176;C and inputted in “Freezer (-20&#176;C) Inventory”) with Leonard lab IBI gel extraction kit, using all iGEM modifications</li>
 +
          <ul>
 +
            <li>sfGFP/mCherry for GG</li>
 +
            <li>Tet-Lrz-GFP</li>
 +
            <li>Cas-Lrz-SS</li>
 +
          </ul>
 +
          <div class="row">
 +
            <div class="col-xs-8"><img src="https://static.igem.org/mediawiki/2016/9/97/T--Northwestern--08_31_1.png" width="1032" height="980" alt=""/></div>
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          </div>
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        </ul>
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      </div>
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    </div>
 +
    <div class="row">
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      <div class="col-sm-2">
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        <p>Tyler</p>
 +
      </div>
 +
      <div class="col-sm-10">
 +
        <ul>
 +
          <li>Gibson Reaction for Ycdo into Lnrzed Cas9</li>
 +
          <ul>
 +
            <li>1.56 µL backbone</li>
 +
            <li>0.29µL insert (Ycdo)</li>
 +
            <li>3.15µL water</li>
 +
            <li>5 µL Gibson mix</li>
 +
          </ul>
 +
          <li>Negative Control</li>
 +
          <ul>
 +
            <li>1.56 µL</li>
 +
            <li>3.44 µL water</li>
 +
            <li>5 µL Gibson mix</li>
 +
          </ul>
 +
          <li>Positive Control</li>
 +
          <ul>
 +
            <li>5µL (+) control</li>
 +
            <li>5µL master mix</li>
 +
          </ul>
 +
          <li>Transformation</li>
 +
          <ul>
 +
            <li>1 µL (+) transformation control (CamR)</li>
 +
            <li>2 µL Gibson (+) control (AmpR)</li>
 +
            <li>2 µL Gibson (-) Control (CamR)</li>
 +
            <li>3 µL Gibson Product (CamR)</li>
 +
            <li>3 µL gRNA Product (TetR) </li>
 +
          </ul>
 +
        </ul>
 +
      </div>
 +
    </div>
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<script type="text/javascript" src="https://2016.igem.org/Team:Northwestern/libraries/jquery?action=raw&ctype=text/javascript"></script>
 
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Revision as of 01:34, 16 October 2016

Notebook

Wednesday, August 31st

Tasks:

Michelle

  • Miniprepped gRNA overnight colony
    • 216.0 ng/uL, 260/280: 1.87, 260/230: 2.20
  • Made nfH2O aliquots
  • Autoclaved things
  • Grew up TetR-Cas9 for western blot
    • 50 mL culture
    • Starting OD600: 0.035 A
    • 1 hour OD600: 0.048 A
    • 2 hour OD600: 0.157 A
    • 2.5 hour OD600: 0.249 A
    • 3 hour OD600: 0.464 A
    • 3.25 hr OD600: 0.600 A

Tasfia

  • Gel extracted PCR products (stored in -20°C and inputted in “Freezer (-20°C) Inventory”) with Leonard lab IBI gel extraction kit, using all iGEM modifications
    • sfGFP/mCherry for GG
    • Tet-Lrz-GFP
    • Cas-Lrz-SS

Tyler

  • Gibson Reaction for Ycdo into Lnrzed Cas9
    • 1.56 µL backbone
    • 0.29µL insert (Ycdo)
    • 3.15µL water
    • 5 µL Gibson mix
  • Negative Control
    • 1.56 µL
    • 3.44 µL water
    • 5 µL Gibson mix
  • Positive Control
    • 5µL (+) control
    • 5µL master mix
  • Transformation
    • 1 µL (+) transformation control (CamR)
    • 2 µL Gibson (+) control (AmpR)
    • 2 µL Gibson (-) Control (CamR)
    • 3 µL Gibson Product (CamR)
    • 3 µL gRNA Product (TetR)