Purify large fragment from Agarose gel >10 Kb

brett brett at BORCIM.WUSTL.EDU
Tue Oct 21 09:03:41 EST 1997

>Hi, Netters,
>Does anybody out there any give me some references or any methods, kits, 
>which work well for purification large fragment from agarose gel?(>10 kB)
>We met unstable results by using Geneclean or Qiagen kit.

Just yesterday I compared three methods for isolating ~200ng of a linear
12kbp fragment from LMT.  I got quantitative recovery by phenol extracting
2x (use no CHCl3) and EtOH pptn of the melted gel (glycogen as carrier).
I tried Boehringer's Agarase followed by pptn, also quantitative.  And I
also extracted the fragment using a homemade "Wizard" miniprep kit (ala
this newsgroup), which gave me ~40% recovery but was the fastest method.

Brett Lindenbach
Washington University - St Louis                  
brett at borcim.wustl.edu                             

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