Difference between revisions of "Team:Austin UTexas LASA/Experiments"

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                         <a data-toggle="collapse" data-parent="#accordion" href="#collapseOne" aria-expanded="true" aria-controls="collapseOne">
 
                         <a data-toggle="collapse" data-parent="#accordion" href="#collapseOne" aria-expanded="true" aria-controls="collapseOne">
 
                             <i class="glyphicon glyphicon-plus"></i>
 
                             <i class="glyphicon glyphicon-plus"></i>
−
                            Protocol Title 1
+
                          PCR (Polymerase Chain Reaction)
 
                         </a>
 
                         </a>
 
                     </h4>
 
                     </h4>
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                 <div id="collapseOne" class="panel-collapse collapse" role="tabpanel" aria-labelledby="headingOne">
 
                 <div id="collapseOne" class="panel-collapse collapse" role="tabpanel" aria-labelledby="headingOne">
 
                     <div class="panel-body animated zoomOut">
 
                     <div class="panel-body animated zoomOut">
−
                        Anim pariatur cliche reprehenderit, enim eiusmod high life accusamus terry richardson ad squid. 3 wolf moon officia aute, non cupidatat skateboard dolor brunch. Food truck quinoa nesciunt laborum eiusmod. Brunch 3 wolf moon tempor, sunt aliqua put a bird on it squid single-origin coffee nulla assumenda shoreditch et. Nihil anim keffiyeh helvetica, craft beer labore wes anderson cred nesciunt sapiente ea proident. Ad vegan excepteur butcher vice lomo. Leggings occaecat craft beer farm-to-table, raw denim aesthetic synth nesciunt you probably haven't heard of them accusamus labore sustainable VHS.
+
<h2>Reactants</h2>
 +
<table>
 +
  <tr>
 +
    <th>10x Reaction Mix</th>
 +
    <th>5 uL</th>
 +
  </tr>
 +
  <tr>
 +
    <th>For primer (10 uM)</th>
 +
    <th>1 uL</th>
 +
  </tr>
 +
  <tr>
 +
    <th>Rev primer (10 uM)</th>
 +
    <th>1 uL</th>
 +
  </tr>
 +
  <tr>
 +
    <th>Template</th>
 +
    <th>1 uL</th>
 +
  </tr>
 +
  <tr>
 +
    <th>Taq Polymerase</th>
 +
    <th>1 uL</th>
 +
  </tr>
 +
  <tr>
 +
    <th>MgCl2 (25 mM)</th>
 +
    <th>1 uL</th>
 +
  </tr>
 +
  <tr>
 +
    <th>DMSO</th>
 +
    <th>1 uL</th>
 +
  </tr>
 +
  <tr>
 +
    <th>NF H2o</th>
 +
    <th>41 uL</th>
 +
  </tr>
 +
  <tr>
 +
    <th><strong>Total</strong></th>
 +
    <th><strong>50 uL</strong></th>
 +
  </tr>
 +
</table>
 +
<h6>Protocol</h6>
 +
<ol>
 +
<li>On ice, add all reagents to PCR tube, Taq polymerase last. </li>
 +
<li>Run thermocycler program:</li>
 +
<ul>
 +
<li>95 C  for  5 min  (2 min if NOT using Hotstart)
 +
</li>
 +
</ul>
 +
</ol>
 +
<p>> 10 cycles:<br>
 +
95 C  for  20 s<br>
 +
0.3 C/s to 50 C<br>
 +
72 C  for  # s (extension = 1 min/kb)<br>
 +
</p>
 +
<p>
 +
> 15 cycles:
 +
<br>95 C  for  20 s                                   
 +
<br>55 C  for  20 s
 +
<br>72 C  for  1 min
 +
</p>
 +
<p>
 +
<br>72 C  for  10 min
 +
<br>4 C  forever
 +
</p>
 
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                 </div>
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                             <i class="glyphicon glyphicon-plus"></i>
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                             Protocol Title 2
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                             PCR Clean Up
 
                         </a>
 
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                 <div id="collapseTwo" class="panel-collapse collapse" role="tabpanel" aria-labelledby="headingTwo">
 
                     <div class="panel-body animated zoomOut">
 
                     <div class="panel-body animated zoomOut">
 +
<table>
 +
  <tr>
 +
    <th>10x Reaction Mix</th>
 +
    <th>5 uL</th>
 +
  </tr>
 +
</table>
 
                         Anim pariatur cliche reprehenderit, enim eiusmod high life accusamus terry richardson ad squid. 3 wolf moon officia aute, non cupidatat skateboard dolor brunch. Food truck quinoa nesciunt laborum eiusmod. Brunch 3 wolf moon tempor, sunt aliqua put a bird on it squid single-origin coffee nulla assumenda shoreditch et. Nihil anim keffiyeh helvetica, craft beer labore wes anderson cred nesciunt sapiente ea proident. Ad vegan excepteur butcher vice lomo. Leggings occaecat craft beer farm-to-table, raw denim aesthetic synth nesciunt you probably haven't heard of them accusamus labore sustainable VHS.
 
                         Anim pariatur cliche reprehenderit, enim eiusmod high life accusamus terry richardson ad squid. 3 wolf moon officia aute, non cupidatat skateboard dolor brunch. Food truck quinoa nesciunt laborum eiusmod. Brunch 3 wolf moon tempor, sunt aliqua put a bird on it squid single-origin coffee nulla assumenda shoreditch et. Nihil anim keffiyeh helvetica, craft beer labore wes anderson cred nesciunt sapiente ea proident. Ad vegan excepteur butcher vice lomo. Leggings occaecat craft beer farm-to-table, raw denim aesthetic synth nesciunt you probably haven't heard of them accusamus labore sustainable VHS.
 
                     </div>
 
                     </div>

Revision as of 22:20, 31 October 2017

Experiments

Reactants

10x Reaction Mix 5 uL
For primer (10 uM) 1 uL
Rev primer (10 uM) 1 uL
Template 1 uL
Taq Polymerase 1 uL
MgCl2 (25 mM) 1 uL
DMSO 1 uL
NF H2o 41 uL
Total 50 uL
Protocol
  1. On ice, add all reagents to PCR tube, Taq polymerase last.
  2. Run thermocycler program:
    • 95 C for 5 min (2 min if NOT using Hotstart)

> 10 cycles:
95 C for 20 s
0.3 C/s to 50 C
72 C for # s (extension = 1 min/kb)

> 15 cycles:
95 C for 20 s
55 C for 20 s
72 C for 1 min


72 C for 10 min
4 C forever

10x Reaction Mix 5 uL
Anim pariatur cliche reprehenderit, enim eiusmod high life accusamus terry richardson ad squid. 3 wolf moon officia aute, non cupidatat skateboard dolor brunch. Food truck quinoa nesciunt laborum eiusmod. Brunch 3 wolf moon tempor, sunt aliqua put a bird on it squid single-origin coffee nulla assumenda shoreditch et. Nihil anim keffiyeh helvetica, craft beer labore wes anderson cred nesciunt sapiente ea proident. Ad vegan excepteur butcher vice lomo. Leggings occaecat craft beer farm-to-table, raw denim aesthetic synth nesciunt you probably haven't heard of them accusamus labore sustainable VHS.
Anim pariatur cliche reprehenderit, enim eiusmod high life accusamus terry richardson ad squid. 3 wolf moon officia aute, non cupidatat skateboard dolor brunch. Food truck quinoa nesciunt laborum eiusmod. Brunch 3 wolf moon tempor, sunt aliqua put a bird on it squid single-origin coffee nulla assumenda shoreditch et. Nihil anim keffiyeh helvetica, craft beer labore wes anderson cred nesciunt sapiente ea proident. Ad vegan excepteur butcher vice lomo. Leggings occaecat craft beer farm-to-table, raw denim aesthetic synth nesciunt you probably haven't heard of them accusamus labore sustainable VHS.
Anim pariatur cliche reprehenderit, enim eiusmod high life accusamus terry richardson ad squid. 3 wolf moon officia aute, non cupidatat skateboard dolor brunch. Food truck quinoa nesciunt laborum eiusmod. Brunch 3 wolf moon tempor, sunt aliqua put a bird on it squid single-origin coffee nulla assumenda shoreditch et. Nihil anim keffiyeh helvetica, craft beer labore wes anderson cred nesciunt sapiente ea proident. Ad vegan excepteur butcher vice lomo. Leggings occaecat craft beer farm-to-table, raw denim aesthetic synth nesciunt you probably haven't heard of them accusamus labore sustainable VHS.
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