High G/C PCR

Nick Jacobsen jacobsen at cf.ac.uk
Fri Oct 25 08:44:49 EST 1996


Hi,
   I have had problems with a high GC PCR. I got it working by doing the 
following. Design your primers with high Tms (round about 78degrees C). 
Get hold of a highly thermostable enzyme (NEB do an enzyme called Deep 
Vent exo-). Set up your rxns as normal and cycle as follows 98C for 1min, 
anneal 1min and do 30 cycles of this. Taq will not survive 98C but Deep 
Vent will. Consequently you get high yield PCR from your GC rich 
template.

hope this helps

cheers

Nick.




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