PCR stutters through oligo-A regions

vilimf01 at mcrcr6.med.nyu.edu vilimf01 at mcrcr6.med.nyu.edu
Mon Mar 18 17:33:26 EST 1996


A recent paper by Chevet et al NAR 23(16):3343-3344 describes the use
of TMACl (60mM) in PCR with A/T rich oligo's.  This method may work
with A/T rich templates as well.  May be worth a look.

		Usual Disclaimers		SVEN





In article <4ik8cg$ju0 at majestix.uni-muenster.de>, Juergen Brosius <RNA.world at uni-muenster.de> writes:
> All polymerases we tried so far (Taq, Tth, Pfu, Vent) seem to have 
> trouble to get faithfully through regions of about 15 A-residues or 
> longer.  The products have usually more As (it varies from clone to 
> clone) than the genomic sequence isolated by regular cloning.  Direct 
> sequencing did not help either.  The region in question reflects the 
> heterogeneity. I presume there are similar problems with other 
> homo-oligomers.  A frightening scenario would be it that behaviour would 
> also persist through oligo-dimers or -trimers.
> Any remedies?
> 
> Thanks in advance.
> Juergen Brosius
> University of Muenster
> e-mail:   RNA.world at uni-muenster.de
> 
>  
> 
> 



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