Q: Inaccuracies in PCR sequencing and genetic variability
ricksonf at bcc.orst.edu
Fri Feb 14 19:45:37 EST 1997
On Thu, 13 Feb 1997, Daniel GAUTHERET wrote:
> I bet this issue has already been debated, but I can't find any relevant
> in the litterature. So here it is:
> Direct sequencing of PCR products often yields sequences with many
> inaccuracies (Ns). Can we say that the position of these undefined
> nucleotides is related to actual genetic variation at these particular
> Thank you.
> Daniel Gautheret
I suspect that if you are getting more than an acceptable amount of
misreads, then you are going to have to clone what must be a
heterogeneous target area. On a "clean" DNA region I can sequence 5
times with maybe two changes each time over 300 bases.
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