PCR mistakes

Clemens Suter-Crazzolara, PhD un691cs at genius.embnet.dkfz-heidelberg.de
Wed Jun 3 04:39:04 EST 1998


Hello,

I am trying to clone a PCR product of 750bp, but it appears to
contain about 4 PCR mistakes. The polymerase I use is Ultma from 
Perkin Elmer. 
As the RNA is of pretty low abundance, I have to amplify for 32 
cycles, still this wasn't much of a problem in the past.

Do you have any idea how to circumvent this high failure rate of
the PCR ? (I re-call lowering the MgCL2 concentration, currently
at 2.5 mM).

Thanks, clemens




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