extracting dna from acrylamide gels
liz_parks at ncsu.edu
Thu Nov 2 11:07:19 EST 2000
Acrylamide will inhibit PCR, EtBr will not. I have had a great deal of
success using a method from Biotechniques (Nov. 1994 Vol. 17, No. 5, pgs
914-921). Basically, cut out the band of interest from the dried or fresh
gel. Incubate it at 95C for 20 minutes in 100 ul of 10 mM Tris-HCl, 50 mM
KCl, 1.5 mM MgCl2, 0.1 % Triton-X, pH 9.0. Use 1 ul in your subsequent PCR
reaction. In my experience, this works better than water.
Jeffrey Markert wrote:
> I'm looking for advice on extracting DNA bands from acrylamide gels. My
> goal is to recover enough DNA for a few PCR reactions. The acrylamide gels
> are stained in EtBr and were made with FMC's Long Ranger mix.
> I've been using the ammonium acetate elution protocol from Maniatis, but
> I'm not finding it to be very reliable in my hands and it would be nice to
> find a procedure that doesn't require an o/n incubation.
> I'm wondering whether there's a simpler and quicker way that will work for
> PCR. Do acrylamide or EtBr inhibit PCR? Could I just elute the gel slice
> in water to avoid having to get rid of the ammonium acetate? Could I just
> freeze (or boil) the gel slice? Any thoughts or anecdotes are welcome.
North Carolina State University
Department of Plant Pathology
Raleigh, NC 27695
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