Team:University College London/Protocols/PCR

From 2012.igem.org

(Difference between revisions)
(Created page with "<noinclude>{{:Team:University_College_London/templates/head|coverpicture=week4}} == Transformation Protocol 2 == </noinclude> '''Step 1 - Setting up PCR tubes:''' Thaw reagents...")
(Transformation Protocol 2)
Line 45: Line 45:
-
{{:Team:University_College_London/templates/pictureinsert|title=Picture 1|url=images/a/af/Ucligem2012Thermo.png}}
 
<noinclude>
<noinclude>
{{:Team:University_College_London/templates/foot}}
{{:Team:University_College_London/templates/foot}}
</noinclude>
</noinclude>

Revision as of 09:18, 22 August 2012

Transformation Protocol 2

Step 1 - Setting up PCR tubes: Thaw reagents and add to PCR tubes in the proportions described in the table below

PCR Components Volume (ul)
5x Reaction Buffer 10
25mM MgCl2 4
10mM dNTPs 1
10uM Forward primer 5
10uM Reverse primer 5
DNA Polymerase 0.25
Nuclease Free Water 24.25
Template DNA 0.5
Total Volume 0.5

Step 2 - PCR program: Add PCR tubes to a thermocycler and run under the following conditions.

PCR conditions Temp (oC) Time (s)
Initial Denaturation (1 cycle) 95 30
Denaturation/Annealing/Extension (30 cycles) 95/55/72 10/25/120
Final Extension (1 cycle) 72 600
Hold 4