PCR problem

Dr G R Taylor ai80 at dial.pipex.com
Sun Jun 9 17:51:49 EST 1996


stephane angers wrote:
> 
> We are trying to amplify a Dna sequence with a lot of CG and after Pcr
> we can't get any amplification (no band on gel) we thing that the
> polymerase is blocked in some secondary structures that are formed.
> Is there  a way to counter that ?
> Stephane Angers
> sangers at accent.net

Stephan:

The following may help:  1) Use deazadGTP:dGTP at 3:1
			 2) Include 10% DMSO
			 3) Increase (double) concentration of polymerase

Look into the literature on Fragile X amplification,  which is very G+C 
rich.

Good luck

-- 
>From GR Taylor PhD MRCPath 
Molecular Genetics, Leeds, UK
gtaylor at hgmp.mrc.ac.uk
-in vitro veritas-




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