DNA isolation from agarose gels
lacyp at unicorn.it.wsu.edu
Sat Mar 6 17:10:34 EST 1999
Annette C. Hollmann (ah690549 at bcm.tmc.edu) wrote:
: In article <KTZD2.351$6D3.7799136 at newsie.cais.net> "Sean Fitzsimmons" <fitzsp at cais.net> writes:
: >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
: I've had good results with the Qiagen QiaQuick gel extraction kit.
One of things I did with the Qiagen extraction system was to not use
dd-water, but use 10 mM Tris (pH 7.5) for resuspending DNA from the
pellet. After switching to this, my recovery jumped to the 70% range at
least. The distilled water I used originally was around pH 6 or so.
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