Short PCR product

Bruno Cenni bruno.removethis.cenni at removethis.insel.ch
Wed Jan 9 02:22:55 EST 2002


"aes" <ali at image.dk> wrote in message news:a1egi1$aha$1 at news.net.uni-c.dk...
> I am going to attempt to do quantitative PCR with degenerate primers, I
can
> make the primers so that they should amplify all the genes I want, but
again
> the primers end up being 28 bp long with 8 bases (in the middle) having
> degenerate composition (R,K and Y). Do you think it is possible to get
these
> primers working in a semi quantitative experiment?
>
> Thank you
>
> Aes

Hi Ali,

I agree with the others that accurate quantification is probably impossible.
But it might depend on the level of accuracy you want, I just saw a
reference in which they used degenerate RT-PCR for differential display of
zinc finger motifs (Anal. Biochem. 1996 236(2):348-52)... so it might depend
on the primer set and how specific they are.

Cheers

Bruno





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