Team:Groningen/protocols/PCR

From 2013.igem.org

(Difference between revisions)
Line 27: Line 27:
<ul type="square">
<ul type="square">
<li>PCR tubes</li>
<li>PCR tubes</li>
-
<li>F- and R-primer</li>
 
-
<li>DNTPs</li>
 
-
<li>Phusion buffer</li>
 
<li>MQ water</li>
<li>MQ water</li>
 +
<li>DMSO 5%</li>
 +
<li>Phusion buffer</li>
 +
<li>DNTPs</li>
 +
<li>F- and R-primer</li>
<li>Phusion Polymerase</li>
<li>Phusion Polymerase</li>
<li>DNA template</li>
<li>DNA template</li>
Line 37: Line 38:
<p>
<p>
<br>
<br>
-
<h5>PCR reaction mixture</h5>
+
<h5>PCR reaction mixture:</h5>
<table border="1">
<table border="1">
   <tr>
   <tr>

Revision as of 10:42, 28 July 2013

PCR

Materials:
  • PCR tubes
  • MQ water
  • DMSO 5%
  • Phusion buffer
  • DNTPs
  • F- and R-primer
  • Phusion Polymerase
  • DNA template


PCR reaction mixture:
Component 50µl Final concentration
MQ water up to 50µl
5x Phusion Buffer 10µl 1x
DMSO 5% 1.5µl
10mM dNTPs 1µl 200µM
primer F 2.5µl 0.5µM
primer R 2.5µl 0.5µM
template DNA 1µl ∼1ng
Phusion Polymerase
2U/µl		 0.3µl 0.01U/µl


Cycling instructions:
Temperature Time Number of Cycles
98°C 2min 1td>
98°C 10sec 34
Tm-5°C 25sec
72°C 30sec/kb
72°C 10min 1
4°C ∞ 1

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