PCR problem

bio_hannan at emuvax.emich.edu bio_hannan at emuvax.emich.edu
Mon Jun 10 17:28:21 EST 1996


In article <31BB5585.10A5 at dial.pipex.com>, Dr G R Taylor <ai80 at dial.pipex.com> writes:
> 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-
> 
I have also heard that increased DMSO conc. (like 8%) workd in some cases.

Gary Hannan



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