Digitonin for permeabilization

Jill Johnston johnstoj at acs.ucalgary.ca
Thu Dec 21 16:21:43 EST 2000


We use digitonin to permeabilize sperm heads so we can use the nuclei to
generate transgenic Xenopus.  We have been having problems controlling
the reaction.  After a 5 min exposure to digitonin, a solution
containing 3% BSA is added to quench/stop the reaction.  It looks like
the nuclei are lysing and we get a large gel-like mass that I think is
DNA.  Has anyone used BSA to stop the action of digitonin (or
lysolecithin)?
Thanks

--
Jillian Johnston
Dept. of Biochemistry and Molecular Biology
University of Calgary
Phone (403) 220-3559
Fax   (403) 270-0737







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