rhubner at gins.uia.ac.be
Thu Dec 10 16:13:59 EST 1998
In article <366FB62C.13C30B31 at liv.ac.uk>, "Richard V. Giles"
<giles at liv.ac.uk> wrote:
> Csaba Jeney wrote:
> > We are trying to get a PCR product using a very long primer pair (60bp
> > each, 20 bp anealing length) but until now there is nothing to glad.
> > We already have changed the annealing temperature, the Mg
> > concentration and the next step will be the primer concentration.
> > Could anybody out there suggest other things and/or suggest the ranges
> > of these variables to get of rid this problem.
> > Thank you in advance.
> > Csaba
> My experience with such primers (I use them to introduce phage RNA pol
> sites) is that they behave just like shorter primers (missing the pol
Caution: in case you use a "proofreading" pol or a mix where it is present
at a high rate, your long primer can get nibbled if not protected...
All the best,
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