Difference between revisions of "Team:Northwestern/07 21"

(Created page with "<html> <style> </style> <head> <meta charset="UTF-8"> <meta http-equiv="X-UA-Compatible" content="IE=edge"> <meta name="viewport" content="width=device-width, initial-...")
 
 
(One intermediate revision by the same user not shown)
Line 20: Line 20:
 
   <article>
 
   <article>
 
     <h1>Thursday, July 21<sup>st</sup></h3>
 
     <h1>Thursday, July 21<sup>st</sup></h3>
−
    <h2>Agenda:</h2>
+
<h2>Tasks:</h2>
−
 
+
<div class="row">
−
<footer id="nav">
+
  <div class="col-sm-2">
 +
    <p>Jordan</p>
 +
  </div>
 +
  <div class="col-sm-10">
 +
    <ul>
 +
      <li>Transformed J04550 RFP part to amplify for gRNA construct
 +
        <ul>
 +
          <li>Resuspended Kit Plate 4, well 2H in 10 uL of water, transformed 2 uL of that into 50 uL comp cells</li>
 +
          <li>Plated 50 and 100 uL</li>
 +
        </ul>
 +
      </li>
 +
      <li>Transformed competent cell test
 +
        <ul>
 +
          <li>7 test conditions: 50, 20, 10, 5, and .5 pg/uL of RFP plasmid (part J04550) and 1.11 ng/ul and 8.9 ng/ul of pSB1C3 + tet plasmid</li>
 +
          <li>Transformed 1 ul of each</li>
 +
          <li>Did two replicate plates of each</li>
 +
        </ul>
 +
      </li>
 +
    </ul>
 +
  </div>
 +
</div>
 +
<div class="row">
 +
  <div class="col-sm-2">
 +
    <p>Michelle</p>
 +
  </div>
 +
  <div class="col-sm-10">
 +
    <ul>
 +
      <li>PCR INP part out of the iGEM registry</li>
 +
      <li>10 uL dH<sub>2</sub>O, pipet up and down, sit 5 min</li>
 +
      <li>Transformed INP biobrick</li>
 +
      <ul>
 +
        <li>Transformed using 1 uL of diluted DNA from plate</li>
 +
        <li>Followed bootcamp protocol except:
 +
          <ul>
 +
            <li>50 mL competent cells instead of 100 mL</li>
 +
            <li>450 uL SOC instead of 900 uL</li>
 +
            <li>Used 108.3 ng/uL DNA straight from the iGEM plate</li>
 +
          </ul>
 +
        </li>
 +
        <li>Plated 100 uL</li>
 +
      </ul>
 +
     
 +
      <li>Ran gel on PCR product</li>
 +
    </ul>
 +
  </div>
 +
</div>
 +
<div class="row">
 +
<div class="col-sm-2">
 +
  <p>Paul</p>
 +
</div>
 +
<div class="col-sm-10">
 +
<ul>
 +
<li>Planned gRNA assembly procedure</li>
 +
<li>Selected storage plasmids for use in Golden Gate</li>
 +
<li>Transformed mRFP device:
 +
<ul>
 +
  <li>Transformed using 1 uL of diluted DNA from plate</li>
 +
  <li>Followed bootcamp protocol except:
 +
    <ul>
 +
      <li>50 mL competent cells instead of 100 mL</li>
 +
      <li>450 uL SOC instead of 900 uL</li>
 +
      <li>Used 108.3 ng/uL DNA straight from the iGEM plate</li>
 +
    </ul>
 +
  </li>
 +
  <li>Plated 100 uL</li>
 +
</ul></li>
 +
</ul>
 +
</div>
 +
</div>
 +
<div class="row">
 +
  <div class="col-sm-2">
 +
    <p>Sam</p>
 +
  </div>
 +
  <div class="col-sm-10">
 +
    <ul>
 +
      <li>Went to the products showcase thing to ask for free stuff</li>
 +
      <li>Found more antibiotic experts</li>
 +
    </ul>
 +
  </div>
 +
</div>
 +
<div class="row">
 +
  <div class="col-sm-2">
 +
    <p>Sara</p>
 +
  </div>
 +
  <div class="col-sm-10">
 +
    <ul>
 +
      <li>Nanodropped the Gibson Tet Cas9 product - 1730 ng/uL</li>
 +
      <li>Diluted the Gibson product to 50 ng/uL
 +
        <ul>
 +
          <li>1.16 uL Gibson product</li>
 +
          <li>38.84 uL NF water</li>
 +
          <li>40 uL in total</li>
 +
        </ul>
 +
      </li>
 +
      <li>Restriction digestion of Gibson
 +
        <ul>
 +
          <li>5 uL 10X NE buffer 4</li>
 +
          <li>1 uL EcoRi HF</li>
 +
          <li>20 uL 50 ng/uL Diluted Gibson (1 ug)</li>
 +
          <li>25 uL water</li>
 +
          <li>50 uL in total</li>
 +
        </ul>
 +
      </li>
 +
      <li>Ran a gel of the restriction digest with Shu
 +
        <ul>
 +
          <li>8 uL ladder</li>
 +
          <li>6 uL DNA
 +
            <ul>
 +
              <li>2.5 uL restricted DNA (50 ng)</li>
 +
              <li>2.5 uL NF water</li>
 +
              <li>1 uL loading dye</li>
 +
            </ul>
 +
          </li>
 +
        </ul>
 +
      </li>
 +
    </ul>
 +
  </div>
 +
</div>
 +
<div class="row">
 +
  <div class="col-sm-2">
 +
    <p>Shu</p>
 +
  </div>
 +
  <div class="col-sm-10">
 +
    <ul>
 +
      <li>Ran a gel of the restriction digest with Sara</li>
 +
      <li>Sorting out the things we received from IDT today</li>
 +
      <li>Read Golden Gate papers</li>
 +
      <li>Looked into the traveling grant</li>
 +
    </ul>
 +
  </div>
 +
</div>
 +
<div class="row">
 +
  <div class="col-sm-2">
 +
    <p>Tasfia</p>
 +
  </div>
 +
  <div class="col-sm-10">
 +
    <ul>
 +
    <li>PCR’ed INP and ran a gel for product
 +
      <ul>
 +
        <li>2.5 uL 10X PCR buffer</li>
 +
        <li>0.5 uL 10 mM dNTPs</li>
 +
        <li>0.5 uL INP-fwd at 10 uM</li>
 +
        <li>0.5 uL INP-rev at 10 uM</li>
 +
        <li>0.5 uL template (BB part, 2.4 kb, so 2 min 24s of elongation)</li>
 +
        <li>1 hr at 50oC Total of 20 uL</li>
 +
      </ul>
 +
      </li>
 +
      <li>Transformed INP</li>
 +
      <ul>
 +
        <li>Transformed using 1 uL of diluted DNA from plate</li>
 +
        <li>Followed bootcamp protocol except:
 +
          <ul>
 +
            <li>50 mL competent cells instead of 100 mL</li>
 +
            <li>450 uL SOC instead of 900 uL</li>
 +
            <li>Used 108.3 ng/uL DNA straight from the iGEM plate</li>
 +
          </ul>
 +
        </li>
 +
        <li>Plated 100 uL</li>
 +
      </ul>
 +
      <li>Worked on presentation slideshow</li>
 +
    </ul>
 +
  </div>
 +
</div>
 +
</article>
 +
  <footer id="nav">
 
       <div class="row">
 
       <div class="row">
 
         <div class="col-sm-4"><a href="https://2016.igem.org/Team:Northwestern/07_20"><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/07_20"><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/07_22">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/07_22">tomorrow <img src="https://static.igem.org/mediawiki/2016/6/65/T--Northwestern--forwardarrow.png" height="15" width="15"/></a></div>
 
       </div>
 
       </div>
 
     </footer>
 
     </footer>
−
  </article>
 
 
<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>

Latest revision as of 15:27, 18 October 2016

Notebook

Thursday, July 21st

Tasks:

Jordan

  • Transformed J04550 RFP part to amplify for gRNA construct
    • Resuspended Kit Plate 4, well 2H in 10 uL of water, transformed 2 uL of that into 50 uL comp cells
    • Plated 50 and 100 uL
  • Transformed competent cell test
    • 7 test conditions: 50, 20, 10, 5, and .5 pg/uL of RFP plasmid (part J04550) and 1.11 ng/ul and 8.9 ng/ul of pSB1C3 + tet plasmid
    • Transformed 1 ul of each
    • Did two replicate plates of each

Michelle

  • PCR INP part out of the iGEM registry
  • 10 uL dH2O, pipet up and down, sit 5 min
  • Transformed INP biobrick
    • Transformed using 1 uL of diluted DNA from plate
    • Followed bootcamp protocol except:
      • 50 mL competent cells instead of 100 mL
      • 450 uL SOC instead of 900 uL
      • Used 108.3 ng/uL DNA straight from the iGEM plate
    • Plated 100 uL
  • Ran gel on PCR product

Paul

  • Planned gRNA assembly procedure
  • Selected storage plasmids for use in Golden Gate
  • Transformed mRFP device:
    • Transformed using 1 uL of diluted DNA from plate
    • Followed bootcamp protocol except:
      • 50 mL competent cells instead of 100 mL
      • 450 uL SOC instead of 900 uL
      • Used 108.3 ng/uL DNA straight from the iGEM plate
    • Plated 100 uL

Sam

  • Went to the products showcase thing to ask for free stuff
  • Found more antibiotic experts

Sara

  • Nanodropped the Gibson Tet Cas9 product - 1730 ng/uL
  • Diluted the Gibson product to 50 ng/uL
    • 1.16 uL Gibson product
    • 38.84 uL NF water
    • 40 uL in total
  • Restriction digestion of Gibson
    • 5 uL 10X NE buffer 4
    • 1 uL EcoRi HF
    • 20 uL 50 ng/uL Diluted Gibson (1 ug)
    • 25 uL water
    • 50 uL in total
  • Ran a gel of the restriction digest with Shu
    • 8 uL ladder
    • 6 uL DNA
      • 2.5 uL restricted DNA (50 ng)
      • 2.5 uL NF water
      • 1 uL loading dye

Shu

  • Ran a gel of the restriction digest with Sara
  • Sorting out the things we received from IDT today
  • Read Golden Gate papers
  • Looked into the traveling grant

Tasfia

  • PCR’ed INP and ran a gel for product
    • 2.5 uL 10X PCR buffer
    • 0.5 uL 10 mM dNTPs
    • 0.5 uL INP-fwd at 10 uM
    • 0.5 uL INP-rev at 10 uM
    • 0.5 uL template (BB part, 2.4 kb, so 2 min 24s of elongation)
    • 1 hr at 50oC Total of 20 uL
  • Transformed INP
    • Transformed using 1 uL of diluted DNA from plate
    • Followed bootcamp protocol except:
      • 50 mL competent cells instead of 100 mL
      • 450 uL SOC instead of 900 uL
      • Used 108.3 ng/uL DNA straight from the iGEM plate
    • Plated 100 uL
  • Worked on presentation slideshow