Team:Groningen/protocols/PCR

From 2013.igem.org

(Difference between revisions)
Line 36: Line 36:
<br>Steps:
<br>Steps:
-
 
+
<table border = "1">
 +
<tr><td>Component</td>
 +
<td>50&#956;l reaction</td>
 +
<td>Final concentration</td>
 +
</tr>
 +
<tr><td>H<sub>2</sub>O</td>
 +
<td>up to 50&#956;l</td>
 +
<td></td>
 +
</tr>
 +
<tr><td>5x Phusion Buffer</td>
 +
<td>10</td>
 +
<td>1x</td>
 +
</tr>
 +
<tr><td>10mM dNTPs</td>
 +
<td>1</td>
 +
<td>0.2&#956;M</td>
 +
</tr>
 +
<tr><td>F-primer</td>
 +
<td></td>
 +
<td>0.2&#956;M</td>
 +
</tr>
 +
<tr><td>R-primer</td>
 +
<td></td>
 +
<td>0.2&#956;M</td>
 +
</tr>
 +
<tr><td>Template DNA</td>
 +
<td>1&#956;l</td>
 +
<td>~1ng</td>
 +
</tr>
 +
<tr><td>Phusion Pol.</td>
 +
<td>0.5&#956;l</td>
 +
<td>0.02U/&#956;l</td>
 +
</tr>
 +
</table>
<br>Cycling instructions:
<br>Cycling instructions:

Revision as of 09:08, 26 July 2013

PCR


Materials:
  • PCR tubes
  • F- and R-primer
  • DNTPs
  • Phusion buffer
  • H2O
  • Phusion Polymerase
  • DNA template

Steps:
Component 50μl reaction Final concentration
H2O up to 50μl
5x Phusion Buffer 10 1x
10mM dNTPs 1 0.2μM
F-primer 0.2μM
R-primer 0.2μM
Template DNA 1μl ~1ng
Phusion Pol. 0.5μl 0.02U/μl

Cycling instructions:
Reference: http://www.thermoscientificbio.com/uploadedFiles/Resources/f-530s-product-information.pdf