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
Sean, I have tried several methods to extract DNA fragments from agarose
(including BM Agarose Gel DNA Extraction Kit). However, I am most pleased
with the yield and quality of the isolated DNA with a low-melting-temperature
agarose extraction. You can find the protocol in the Maniatus Molecular
Cloning manual on page 6.30. Feel free to email me with any questions.
Josh Neufeld, Graduate Student
Macdonald Campus of Mcgill University/
Pulp and Paper Research Institute of Canada
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