Problem Blotting of Pulsed Field Gels
J. Pat Martinez
martinez at puccini.CRL.umn.edu
Tue Dec 17 11:01:39 EST 1996
We are currently having difficulty blotting Soybean high mol. weight
DNA. In the past, about 1 year ago, everything worked fine, its only
recently that we've had problems. Even the Lambda standards are faint on
the probed filters. We've tested the following to no avail.
Depurination in 0.25M HCL for 0,10, or 20 minutes
Neutral vs. alkaline blotting
Traditional capillary blotting (24 hrs.) vs. vacuum blotting
1% 'standard' agarose vs. 1% Fast Lane agarose
We use Whatman 3M filter paper and Hybond N+ membranes. Any suggestions
to improve our blotting of PFGE gels?
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