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:
> Hi,
> 
>    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

-- 
Richard P. Grant MA DPhil      rpgrant at molbiol.ox.ac.uk
Nuffield Department of Obstetrics and Gynaecology, University of Oxford.
http://users.ox.ac.uk/~lady0266
No wonder I can't go to parties anymore.



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