PCR Puzzler

Rudi van de Wetering ReplyToButNoSpamR.vandewetering at czzorlnm.azn.nl
Fri Nov 17 11:23:18 EST 2000


> I am amplifying a peptide receptor in rat prostate and typically get the
> predicted product of 500 bp. However, there is usually a minor band at
> 600 bp. I never gave this much thought since there is a possible false
> priming site, which would give a minor product at that size. However,
> the other week I surveyed several tissues and discovered that in some
> tissues this minor product was absent or greatly reduced while in one
> tissue (spleen)  the minor product was much more prevalent than the 500
> bp product.
>
> Great, I thought ? alternate splicing or some related but new receptor.
> However, when I gel purified the 600 bp band from spleen and then
> reamplified it, what I got was a major product at 500 bp and a minor
> product at 600 bp. So it looks like false priming again. But why would I
> see tissue specificity for false priming and the minor product?
>
> Any ideas from you PCR aficionados out there?

Have you sequenced both products to see what they really are? That would
help before we start of speculating.

Rudi







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