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

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   <article>
 
   <article>
 
     <h1>Monday, August 8<sup>th</sup></h3>
 
     <h1>Monday, August 8<sup>th</sup></h3>
−
     <h2>Agenda:</h2>
+
     <h2>Tasks:</h2>
−
 
+
    <div class="row">
 +
      <div class="col-sm-2">
 +
        <p>Sara, Jordan, Shu, Tasfia</p>
 +
      </div>
 +
      <div class="col-sm-10">
 +
        <ul>
 +
          <li>Ran Golden Gate reaction, transformed 5uL of each reaction</li>
 +
        </ul>
 +
        <table width="100%" border="0" cellpadding="2px">
 +
          <tbody>
 +
            <tr>
 +
              <th scope="col">mCherry1</th>
 +
              <th scope="col">GFP1-mCherry2</th>
 +
              <th scope="col">GFP1-GFP2-mCherry3</th>
 +
              <th scope="col">GFP1-GFP2-GFP3-mCherry4</th>
 +
              <th scope="col">GFP1-GFP2-GFP3-GFP4-mCherry5</th>
 +
            </tr>
 +
            <tr>
 +
              <td><ul>
 +
                  <li>2 uL BB</li>
 +
                  <li>2 uL 10X Ligase buffer</li>
 +
                  <li>1 uL ligase</li>
 +
                  <li>0.5 uL BsaI</li>
 +
                  <li>1 uL SS</li>
 +
                  <li>1 uL mC1</li>
 +
                  <li>12.5 uL H<sub>2</sub>0</li>
 +
                </ul></td>
 +
              <td><ul>
 +
                  <li>2 uL BB</li>
 +
                  <li>2 uL 10X Ligase buffer</li>
 +
                  <li>1 uL ligase</li>
 +
                  <li>0.5 uL BsaI</li>
 +
                  <li>1 uL SS</li>
 +
                  <li>1 uL G1</li>
 +
                  <li>1 uL mC2</li>
 +
                  <li>11.5 uL H<sub>2</sub>0</li>
 +
                </ul></td>
 +
              <td><ul>
 +
                  <li>2 uL BB</li>
 +
                  <li>2 uL 10X Ligase buffer</li>
 +
                  <li>1 uL ligase</li>
 +
                  <li>0.5 uL BsaI</li>
 +
                  <li>1 uL SS</li>
 +
                  <li>1 uL G1</li>
 +
                  <li>1 ul G2</li>
 +
                  <li>1 uL mC3</li>
 +
                  <li>10.5 uL H<sub>2</sub>0</li>
 +
                </ul></td>
 +
              <td><ul>
 +
                  <li>2 uL BB</li>
 +
                  <li>2 uL 10X Ligase buffer</li>
 +
                  <li>1 uL ligase</li>
 +
                  <li>0.5 uL BsaI</li>
 +
                  <li>1 uL SS</li>
 +
                  <li>1 uL G1</li>
 +
                  <li>1 ul G2</li>
 +
                  <li>1 uL G3</li>
 +
                  <li>1 uL mC4</li>
 +
                  <li>9.5 uL H<sub>2</sub>0</li>
 +
                </ul></td>
 +
              <td><ul>
 +
                  <li>2 uL BB</li>
 +
                  <li>2 uL 10X Ligase buffer</li>
 +
                  <li>1 uL ligase</li>
 +
                  <li>0.5 uL BsaI</li>
 +
                  <li>1 uL SS</li>
 +
                  <li>1 uL G1</li>
 +
                  <li>1 ul G2</li>
 +
                  <li>1 uL G3</li>
 +
                  <li>1 ul G4</li>
 +
                  <li>1 uL mC5</li>
 +
                  <li>8.5 uL H<sub>2</sub>0</li>
 +
                </ul></td>
 +
            </tr>
 +
          </tbody>
 +
        </table>
 +
      </div>
 +
    </div>
 +
    <div class="row">
 +
      <div class="col-sm-2">
 +
        <p>Paul</p>
 +
      </div>
 +
      <div class="col-sm-10">
 +
        <ul>
 +
          <li>Submitted sequencing order </li>
 +
          <li> Ran gel salt test (salt+ diH2O *not autoclaved)
 +
            <ul>
 +
              <li>Added 1 uL of salt solution to mixture:
 +
                <ul>
 +
                  <li>Ladder:
 +
                    <ul>
 +
                      <li>2 uL ladder</li>
 +
                      <li>6 uL blue dye</li>
 +
                      <li>1 uL salt solution</li>
 +
                    </ul>
 +
                  </li>
 +
                  <li>Tet lin for Cas9 (~2.2 kb):
 +
                    <ul>
 +
                      <li>4 uL DNA</li>
 +
                      <li>1 uL dye</li>
 +
                      <li>1 uL salt solution</li>
 +
                    </ul>
 +
                  </li>
 +
                </ul>
 +
              </li>
 +
              <li>Lanes 1-5 (from left to right): NEB 2-log purple ladder: 50 mM, 10 mM, 1 mM, 0.5 mM, no salt*</li>
 +
              <li>Lanes 6-10: Tet linearized for Cas9: 50 mM, 10 mM, 1 mM, 0.5 mM, no salt*</li>
 +
              <li>Should’ve added 1 uL of diH2O to control for the addition of impure water, but instead added nothing (5 uL DNA+ 1 uL dye)</li>
 +
            </ul>
 +
            <div class="row">
 +
              <div class="col-sm-6"><img src="https://static.igem.org/mediawiki/2016/9/9a/T--Northwestern--08_08_1.png" width="735" height="832" alt=""/></div>
 +
            </div>
 +
          </li>
 +
          <li>Plated high copy Cam resistant cells from Patrick on our Cam plates, incubated</li>
 +
        </ul>
 +
      </div>
 +
    </div>
 +
    <div class="row">
 +
      <div class="col-sm-2">
 +
        <p>Sara</p>
 +
      </div>
 +
      <div class="col-sm-10">
 +
        <ul>
 +
          <li>Made cam plates
 +
            <ul>
 +
              <li>275 mL LB</li>
 +
              <li>4.5 g bacto agar</li>
 +
              <li>Autoclave, then 275 uL Cam</li>
 +
            </ul>
 +
          </li>
 +
          <li>Got 6X purple loading dye from the Jewett lab</li>
 +
        </ul>
 +
      </div>
 +
    </div>
 +
    <div class="row">
 +
      <div class="col-sm-2">
 +
        <p>Shu</p>
 +
      </div>
 +
      <div class="col-sm-10">
 +
        <ul>
 +
          <li>Replate the transformed Gibson products in the fridge</li>
 +
          <li>Retransform the Gibson products in the freezer </li>
 +
        </ul>
 +
      </div>
 +
    </div>
 +
    <div class="row">
 +
      <div class="col-sm-2">
 +
        <p>Tasfia</p>
 +
      </div>
 +
      <div class="col-sm-10">
 +
        <ul>
 +
          <li>Premixed sequencing rxns, “Recipe” based off of 7.26.2016 doc</li>
 +
          <li>Poured culture media for tet plates (3 mL of 5 mg/mL Tet antibiotic) </li>
 +
        </ul>
 +
      </div>
 +
    </div>
 +
    <div class="row">
 +
      <div class="col-sm-2">
 +
        <p>Tyler</p>
 +
      </div>
 +
      <div class="col-sm-10">
 +
        <ul>
 +
          <li>Premixed sequencing rxns</li>
 +
          <li>Ordered mRFP linearizing primer</li>
 +
          <li>Blasted a few sequences for mathematical modeling, started looking into Kanamycin resistance </li>
 +
        </ul>
 +
      </div>
 +
    </div>
 +
  </article>
 
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         <div class="col-sm-4"><a href="https://2016.igem.org/Team:Northwestern/08_07"><img src="https://static.igem.org/mediawiki/2016/c/c6/T--Northwestern--backarrow.png" height="15" width="15"/> yesterday</a></div>
 
         <div class="col-sm-4"><a href="https://2016.igem.org/Team:Northwestern/08_07"><img src="https://static.igem.org/mediawiki/2016/c/c6/T--Northwestern--backarrow.png" height="15" width="15"/> yesterday</a></div>
−
         <div class="col-sm-4"><a href="https://2016.igem.org/Team:Northwestern/Notebook">back to calender </a></div>
+
         <div class="col-sm-4"><a href="https://2016.igem.org/Team:Northwestern/Notebook">back to calendar </a></div>
 
         <div class="col-sm-4"><a href="https://2016.igem.org/Team:Northwestern/08_09">tomorrow <img src="https://static.igem.org/mediawiki/2016/6/65/T--Northwestern--forwardarrow.png" height="15" width="15"/></a></div>
 
         <div class="col-sm-4"><a href="https://2016.igem.org/Team:Northwestern/08_09">tomorrow <img src="https://static.igem.org/mediawiki/2016/6/65/T--Northwestern--forwardarrow.png" height="15" width="15"/></a></div>
 
       </div>
 
       </div>

Latest revision as of 15:30, 18 October 2016

Notebook

Monday, August 8th

Tasks:

Sara, Jordan, Shu, Tasfia

  • Ran Golden Gate reaction, transformed 5uL of each reaction
mCherry1 GFP1-mCherry2 GFP1-GFP2-mCherry3 GFP1-GFP2-GFP3-mCherry4 GFP1-GFP2-GFP3-GFP4-mCherry5
  • 2 uL BB
  • 2 uL 10X Ligase buffer
  • 1 uL ligase
  • 0.5 uL BsaI
  • 1 uL SS
  • 1 uL mC1
  • 12.5 uL H20
  • 2 uL BB
  • 2 uL 10X Ligase buffer
  • 1 uL ligase
  • 0.5 uL BsaI
  • 1 uL SS
  • 1 uL G1
  • 1 uL mC2
  • 11.5 uL H20
  • 2 uL BB
  • 2 uL 10X Ligase buffer
  • 1 uL ligase
  • 0.5 uL BsaI
  • 1 uL SS
  • 1 uL G1
  • 1 ul G2
  • 1 uL mC3
  • 10.5 uL H20
  • 2 uL BB
  • 2 uL 10X Ligase buffer
  • 1 uL ligase
  • 0.5 uL BsaI
  • 1 uL SS
  • 1 uL G1
  • 1 ul G2
  • 1 uL G3
  • 1 uL mC4
  • 9.5 uL H20
  • 2 uL BB
  • 2 uL 10X Ligase buffer
  • 1 uL ligase
  • 0.5 uL BsaI
  • 1 uL SS
  • 1 uL G1
  • 1 ul G2
  • 1 uL G3
  • 1 ul G4
  • 1 uL mC5
  • 8.5 uL H20

Paul

  • Submitted sequencing order
  • Ran gel salt test (salt+ diH2O *not autoclaved)
    • Added 1 uL of salt solution to mixture:
      • Ladder:
        • 2 uL ladder
        • 6 uL blue dye
        • 1 uL salt solution
      • Tet lin for Cas9 (~2.2 kb):
        • 4 uL DNA
        • 1 uL dye
        • 1 uL salt solution
    • Lanes 1-5 (from left to right): NEB 2-log purple ladder: 50 mM, 10 mM, 1 mM, 0.5 mM, no salt*
    • Lanes 6-10: Tet linearized for Cas9: 50 mM, 10 mM, 1 mM, 0.5 mM, no salt*
    • Should’ve added 1 uL of diH2O to control for the addition of impure water, but instead added nothing (5 uL DNA+ 1 uL dye)
  • Plated high copy Cam resistant cells from Patrick on our Cam plates, incubated

Sara

  • Made cam plates
    • 275 mL LB
    • 4.5 g bacto agar
    • Autoclave, then 275 uL Cam
  • Got 6X purple loading dye from the Jewett lab

Shu

  • Replate the transformed Gibson products in the fridge
  • Retransform the Gibson products in the freezer

Tasfia

  • Premixed sequencing rxns, “Recipe” based off of 7.26.2016 doc
  • Poured culture media for tet plates (3 mL of 5 mg/mL Tet antibiotic)

Tyler

  • Premixed sequencing rxns
  • Ordered mRFP linearizing primer
  • Blasted a few sequences for mathematical modeling, started looking into Kanamycin resistance