DNA isolation from agarose gels

jag100 at snethen.com jag100 at snethen.com
Thu Mar 11 17:35:49 EST 1999


In article <KTZD2.351$6D3.7799136 at newsie.cais.net>,
  "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
>
>

I use AgarAce from Promega to extract large (5-10 kb) PCR fragments from
agarose.  This works alright, but could be better.  I would like to try some
of the protocols suggested in this thread, but I am doing this in high
quantity (upwards of 70-80 fragments at a time)--it would be a bit daunting
to prepare individual tubes each time (I am in a 96-well block now.)  Any
ideas for high- throughput (and still high-yield) isolations?

Thanks,

Jen
jag100 at snethen.com

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