DNAse contamination

Hank Seifert h-seifert at nwu.edu
Wed Nov 16 09:28:15 EST 1994


In article <3a6p3u$h2s at noc.usfca.edu>, chihara at noc.usfca.edu (Dr. Chihara) says:
>
>We have been having the same problem. I sure would like to avoid CsCl for 
>such small preps. Has anyone any simpler solutions??Please post the 
>response .Thanks. 

We routinely treat our mini-preps with self-digested pronase (10 ul of a 
10 mg/ml stock in a 300-400 ul boiling prep after spinning out the 'snot')
for 30 min at 65oC. We then precipitate with isopropanol containing 1 mM
PMSF to inactivate the pronase. Fred Heffron worked this out years ago and
it inactivates the nucleases and avoids phenol.

Hank Seifert <h-seifert at nwu.edu>
Department of Microbiology-Immunology
Northwestern University
303 East Chicago Ave.
Chicago, IL 60611
Tel: (312) 503-9788
Fax: (312) 503-9787 or -1339



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