XbaI diestion of PCR products

Raymond Dalgleish ray at le.ac.uk
Fri Jul 23 02:35:43 EST 2004


The SNP definitely IS heterozygous. I know that the digest is incomplete by
density scanning the bands on the gel. There is too much undigested DNA
fragment. It's not due to the formation of heteroduplexes during the late
cycles of the PCR because the same fragment contains a PvuII site which
digests to completion eevn when it too is in the heterozygous state (and on
the same allele as the XbaI site).

Raymond

"Sergio" <yokese at hotmail.com> wrote in message
news:2mao51Fl47tbU1 at uni-berlin.de...
> Dr. Hiranya S. Roychowdhury wrote:
> > XbaI, in my experience, is not one of the finnicky enzymes.  It is
possible
> > that you have been just unfortunate enough to have a misincorporation at
the
> > presumptive XbaI site in the amplicon; isn't it?
>
> or a SNP in heterozygosis??
> Sergio





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