problems with PCR
s_nurrish at icrf.icnet.uk
Wed Jun 22 13:28:53 EST 1994
In article <2u4ed7$lr2 at mserv1.dl.ac.uk>, WMGDDM <WMGDDM at CARDIFF.AC.UK>
> I've got a problem with a PCR that I'm attempting...
> I'm trying to use a pair of primers to follow a series
> of deletions that I'm making in a plasmid.
> Under 'standard' conditions I can amplify the deleted
> version of the plasmid but not the original undeleted
> Can anyone explain this, I've tried all the usual
> variations in the reaction conditions, etc. with no luck!
Is the deleted bit very GC rich, if so this can screw up even Taq.
We've had a lot of problems with particularly GC regions in the 5' end of
the SRF gene. Making the reaction 2.5% formamide got over our problem.
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