"

From 2010.igem.org

Module I, group 2(b)

Using the newly synthesized primers go through a pre-test experiment.

PCR system (FastPFU):

 H2O	        31.4μl
 5×buffer	10μl
 dNTP	        5μl
 primer1	1μl
 primer2	1μl
 template	1μl
 FastPFU	0.6μl
 Total	        50μl

Module I, Group 2c

Conduct PCR to amplify eGFP, Kan and Chlr with DNA polymerase pyrobest.

PCR system (FastPFU):

 H2O	        36.7μl
 10×buffer	5μl
 dNTP	        5μl
 primer1	1μl
 primer2	1μl
 template	1μl
 FastPFU	0.3μl
 Total	        50μl

result

The concentration of the M, K and C are a little lower. We’ll amplify them once again with two degrees lower of the annealing temperature.