IUBio

DNA isolation from agarose gels

John R. McQuiston zje8 at cdc.gov
Tue Mar 9 07:51:34 EST 1999


I have found that using BioRad's buffer and matrix with Promega's columns and 
protocol gives about 90% recovery (under 2kb) from even regular HMP agarose.  
1) cut out band and put in tube with 3vol of BioRad binding buffer.
2) Put at 65C until agarose melts.
3) add 25-50ul of matrix. (based on vol.) Let sit for 2-3 min.
4) follow Promega's protocol from here.  

John



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