freeze-squeeze (Re: geneclean: vortexing)
Jim Graham
graham at biodec.wustl.edu
Mon Aug 5 15:54:52 EST 1996
Just take your EtBr stained low-melt gel slice and put it in the -70 for a
few minutes. Centrifuge it and use the supernatant directly. Works fine for
ligation, sequencing, T7 transcription, T4 blunting, and everything else
I've ever tried. The advantage over Gene-clean, Prep-a-Gene, Quiagen is
that it works EVERY SINGLE TIME, because the visualized DNA cannot be
lost, as only one microcentrifuge tube is involved. :)
Jim
J. Graham PhD
Biology Department
Washington University of St. Louis
>> I have moved to the freeeze-squize long time ago: better yields than
>> GeneClean and no limit in size.
>The problem with freeze-squeeze is the huge elution volume. When you use
>any reasonable amount for a ligation reaction the yields are much lower
>than for GeneClean. Also, you have to use miniscule amounts to avoid
>inhibiting enzymatic reactions. Are my limited experiences with
>freeze-squeeze, which is by far the simplest cleanup method possible
>incorrect?
>Thanks,
>Trevor
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