Team:Sumbawagen/Notebook/protocol2

From 2014.igem.org

(Difference between revisions)
 
(One intermediate revision not shown)
Line 171: Line 171:
             </button>
             </button>
<a class="brand" style="padding:0px 15px;width:70px;height:50px;" href="https://igem.org"><img width="70px" height="50px" src="https://static.igem.org/mediawiki/2013/1/17/IGEM_basic_Logo_white_stylized.png"> </a>
<a class="brand" style="padding:0px 15px;width:70px;height:50px;" href="https://igem.org"><img width="70px" height="50px" src="https://static.igem.org/mediawiki/2013/1/17/IGEM_basic_Logo_white_stylized.png"> </a>
-
             <a class="brand" href="https://2014.igem.org/Team:Sumbawagen/Parts">Sumbawagen</a>
+
             <a class="brand" href="https://2014.igem.org/Team:Sumbawagen">Sumbawagen</a>
             <div class="nav-collapse collapse">
             <div class="nav-collapse collapse">
Line 181: Line 181:
                   <a href="#" class="dropdown-toggle" data-toggle="dropdown">Overviews <b class="caret"></b></a>
                   <a href="#" class="dropdown-toggle" data-toggle="dropdown">Overviews <b class="caret"></b></a>
                   <ul class="dropdown-menu">
                   <ul class="dropdown-menu">
-
                       <li><a href="https://2014.igem.org/Team:Sumbawagen/overviews/Econey"> E-coney Project </a></li>
+
                       <li><a href="https://2014.igem.org/Team:Sumbawagen/overviews/Econey_Project"> Econey Project </a></li>
                     </ul>
                     </ul>
                 </li>
                 </li>
Line 196: Line 196:
<li><a href="https://2014.igem.org/Team:Sumbawagen/parts">Parts</a></li>
<li><a href="https://2014.igem.org/Team:Sumbawagen/parts">Parts</a></li>
<li><a href="https://2014.igem.org/Team:Sumbawagen/Safety">Safety</a></li>  
<li><a href="https://2014.igem.org/Team:Sumbawagen/Safety">Safety</a></li>  
-
<li><a href="https://2014.igem.org/Team:Sumbawagen/Attributions">Attribution</a></li>                          
+
<li><a href="https://2014.igem.org/Team:Sumbawagen/Attributions">Attribution</a></li>
 +
<li><a href="https://2014.igem.org/Team:Sumbawagen/future_direction">Future Direction</a></li>                             
                                
                                
                    
                    
Line 207: Line 208:
                   <a href="#" class="dropdown-toggle" data-toggle="dropdown">Notebook <b class="caret"></b></a>
                   <a href="#" class="dropdown-toggle" data-toggle="dropdown">Notebook <b class="caret"></b></a>
                   <ul class="dropdown-menu">
                   <ul class="dropdown-menu">
-
                     <li><a href="https://2014.igem.org/Team:Sumbawagen/Notebook">Daily Notes</a></li>
+
 
 +
                    <li class="nav-header">Daily Notes</li>
 +
                     <li><a href="https://2014.igem.org/Team:Sumbawagen/Notebook">Policy & Practices</a></li>
 +
                    <li><a href="https://2014.igem.org/Team:Sumbawagen/Notebook2">Lab</a></li>
                     <li><a href="https://2014.igem.org/Team:Sumbawagen/Notebook/Protocol">Protocol</a></li>
                     <li><a href="https://2014.igem.org/Team:Sumbawagen/Notebook/Protocol">Protocol</a></li>
                     <!--<li><a href="#">Improvements</a></li>-->
                     <!--<li><a href="#">Improvements</a></li>-->
Line 219: Line 223:
                   <ul class="dropdown-menu">
                   <ul class="dropdown-menu">
                     <li><a href="https://2014.igem.org/Team:Sumbawagen/Team">Meet the Team</a></li>
                     <li><a href="https://2014.igem.org/Team:Sumbawagen/Team">Meet the Team</a></li>
-
                    <li><a href="http:https://igem.org/Team.cgi">Team Information</a></li>
 
                     <li><a href="https://2014.igem.org/Team:Sumbawagen/Team/Gallery">Gallery</a></li>
                     <li><a href="https://2014.igem.org/Team:Sumbawagen/Team/Gallery">Gallery</a></li>
                     <li><a href="https://2014.igem.org/Team:Sumbawagen/Team/Contact">Contact</a></li>
                     <li><a href="https://2014.igem.org/Team:Sumbawagen/Team/Contact">Contact</a></li>
Line 259: Line 262:
<li><a href="https://2014.igem.org/Team:Sumbawagen/Human_practice/moyo">Moyo Festival</a></li>
<li><a href="https://2014.igem.org/Team:Sumbawagen/Human_practice/moyo">Moyo Festival</a></li>
<li><a href="https://2014.igem.org/Team:Sumbawagen/Human_practice/virtual">Virtual Outreach</a></li>
<li><a href="https://2014.igem.org/Team:Sumbawagen/Human_practice/virtual">Virtual Outreach</a></li>
-
<li><a href="https://2014.igem.org/Team:Sumbawagen/Human_practice/Support">Responds and supports</a></li>
+
<li><a href="https://2014.igem.org/Team:Sumbawagen/Human_practice/Support">Responses and supports</a></li>
                                                            
                                                            

Latest revision as of 09:11, 19 February 2015

Team:Dundee/Team - 2013.igem.org

 

Team:Sumbawagen/Team

From 2014.igem.org

iGEM Sumbawagen 2014 · Econey

Notebook – Protocol – 2.PCR (Polymerase Chain Reaction)

1. Ordered primers were dissolved in 1x TE buffer to get a 100 uM stock, then diluted to 10 uM stock by 1x TE buffer (usually to final volume of 200 ul). Primers stocks were stored at -20 °C.

2. There were 3 pairs of primers for the PCR namely

  • ACEF/ACER1 to clone catalytic domain sequence of adenylate cyclase gene
  • AEF/AER to clone full length sequence of IIA(Glc) gene

3. Reagents compositions for PCR were as follow:

  • 2X PCR master mix (Thermoscientific), 25 ul
  • Sterilized distilled water (Akuabides), 15 ul
  • 10 uM forward primer, 5 ul
  • 10 uM reverse primer, 5 ul
  • Sterilized distilled water (Akuabides), 15 ul
  • 10 uM forward primer, 5 ul
  • DNA template, colony from agar plate 0 ul
  • Total volume, 50 ul

4.Reaction conditions for PCR were as follow:

  • 95 °C, 2 minutes
  • 95 °C, 30 seconds
  • 53 °C, 30 seconds
  • 72 °C, 2 minutes
  • 72 °C, 7 minutes
  • Total cycle, 35 cycles