Difference between revisions of "Team:Gaston Day School/Notebook"

Line 34: Line 34:
 
                     <a role="button" data-toggle="collapse" data-parent="#accordion" href="#P1-collapse" aria-expanded="false" aria-controls="P1-collapse">
 
                     <a role="button" data-toggle="collapse" data-parent="#accordion" href="#P1-collapse" aria-expanded="false" aria-controls="P1-collapse">
 
<div>
 
<div>
−
                     <div class="col-md-11">09/01/2017</div><div class="col-md-1"><i class="fa fa-arrow-down fa-10" aria-hidden="true"></i></div>
+
                     <div class="col-md-11">09/01/2017 PCR Primer</div><div class="col-md-1"><i class="fa fa-arrow-down fa-10" aria-hidden="true"></i></div>
 
</div>
 
</div>
 
                     </a>
 
                     </a>
Line 42: Line 42:
 
                 <div id="P1-collapse" class="panel-collapse collapse" role="tabpanel" aria-labelledby="P1">
 
                 <div id="P1-collapse" class="panel-collapse collapse" role="tabpanel" aria-labelledby="P1">
 
                     <div class="panel-body">
 
                     <div class="panel-body">
−
<p>
 
−
<b>Materials:</b><br>
 
−
LB broth<br>
 
−
Ice<br>
 
−
Selection plates<br>
 
−
<br>
 
−
<b>Methods:</b><br>
 
−
<ol style="font-size:16px;">
 
−
<li>Thaw 50µL competent E. coli cells on ice for 10 minutes<br></li>
 
−
<li>Add:
 
−
<ul style="font-size:16px;">
 
−
<li>5-10 µl DNA from a ligation reaction mix or </li>
 
−
<li>10-100ng DNA of a known plasmid </li>
 
−
</ul>
 
−
</li>
 
−
<li>Carefully flick the tube 4-5 times to mix cells and DNA. <b>Do not vortex.</b></li>
 
−
<li>Place the mixture on ice for 30 minutes. <b>Do not mix.</b></li>
 
−
<li>Heat shock at exactly 42°C for exactly 30 seconds. <b>Do not mix.</b></li>
 
−
<li>Place on ice for 5 minutes. <b>Do not mix.</b></li>
 
−
<li>Pipette 950 µl of room temperature SOC or LB media into the mixture.</li>
 
−
<li>Incubate at 37°C and 200-250 rpm for 60 minutes.</li>
 
−
<li>Mix the cells thoroughly by flicking the tube and inverting.</li>
 
−
<li>Spread:
 
−
<ul style="font-size:16px;">
 
−
<li>For ligation reaction DNA: 100µl of each transformation reaction onto a selection plate. For the rest of 900 µL:
 
−
<ol style="font-size:16px;">
 
−
<li>Pellet cells at 8000rpm for 3 minutes</li>
 
−
<li>Remove and dispense 600 µL of supernatant </li>
 
−
<li>Re-suspend cells by light vortexing</li>
 
−
<li>Plate resuspended cells as above</li>
 
−
</ol></li>
 
  
−
<li>For known plasmid: 10 & 100 µL of each transformation reaction onto a selection plate. For the rest of 890 µL:
+
<table>
−
<ol style="font-size:16px;">
+
    <thead>
−
<li>Pellet cells at 8000rpm for 3 minutes</li>
+
      <tr>
−
<li>Remove and dispense 600 µL of supernatant </li>
+
        <th>14.3</th>
−
<li>Re-suspend cells by light vortexing</li>
+
        <th>H2O</th>
−
<li>Plate resuspended cells as above</li>
+
        <th>114.4</th>
−
</ol></li>
+
      </tr>
−
</ul>
+
    </thead>
−
<li>Incubate overnight at 37°C with plates upside down.</li>
+
    <tbody>
−
</ol>
+
      <tr>
−
 
+
        <th>2.5</th>
−
</p>
+
        <td>buffer</td>
 +
        <td>20</td>
 +
      </tr>
 
     </div>
 
     </div>
 
   </div>
 
   </div>

Revision as of 04:15, 24 October 2017

Notebook

On this page, Gaston Day School iGEM team would like to share with you our notes during the experiments from 09/01/2017 to 10/23/2017. The team leader Heena Saqib records the data patiently.

 

Email: iGEM@gastonday.org

Gaston Day School

2001 Gaston Day School Rd

Gastonia 28056

14.3 H2O 114.4
2.5 buffer 20