Re-PCRing after gel-purification problems
Richard P. Grant
rpgrant at molbiol.ox.ac.uk
Wed Oct 9 10:42:43 EST 1996
In article <barry-0710961116360001 at johnsond3.med.yale.edu>, barry at sccs.swarthmore.edu (Brendan) writes:
> I'm having trouble simply Re-PCRing products after gel purifying them.
> I use TAE gels and SpinX columns to gel purify first round bands. Why is
> this second round not working? Does PCR work with templates that are in
> Acetate (tAe)? Any ideas....I'm stumped.
We routinely use QIAEX to purify from TBE/agarose with no ill effects.
However, given that I'm currently in the midst of a large project, this would
have been expensive for my 2-round PCR expts. So I just cut the bands out of
a gel, melted, took 5 - 10 ul of the agarose and plonked it straight into a 50
ul PCR rxn. Superb.
Richard P. Grant MA DPhil rpgrant at molbiol.ox.ac.uk
Nuffield Department of Obstetrics and Gynaecology, University of Oxford.
No wonder I can't go to parties anymore.
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