DNA isolation from agarose gels

Rick lmll at gpu.srv.ualberta.ca
Wed Mar 10 07:42:04 EST 1999


I'm not sure the method we have been using is teh simplest, but it works fine
to us. We are using the SpinX tube from Corstar(many other company have the
equivalents), just cut you DNA bands (as samll as possible) and spin down for
30 sec. it just takes 1 min (from the time you cut the band to get DNA
solution). Generally there is no need to concentrate by EtOH if you keep your
DNA band small enough, or no need to extract by phenol. It's pure enough for
sequencing, cloning, PCRing...For DNA bands ranging from 0.5 to 3 kb, we
usually get >70% recovered. It's better to us TAE instead of TBE buffer.

Rick




Sean Fitzsimmons <fitzsp at cais.net> wrote:
: What is the simplest and most efficient method for recovering restriction
: fragments from agarose gels.  I am trying to recover PCR product whose ends
: have been digested with restriction enzymes from HMP agarose.  The gels have
: loads of product, but recovery of approximately 5% seems to be the norm.
: I've tried Wizard PCR preps, Quiex II, and electroelution, all with very
: dissappointing yields.  Any suggestions of simple, high-yield protocols?

: Thanks in advance.

: Sean Fitzsimmons





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