genomic southerns

Stillman Stillman at BIOSCIENCE.BIOLOGY.UTAH.EDU
Fri Jul 30 17:13:37 EST 1993


                       Subject:                                Time: 2:42 PM
  OFFICE MEMO          genomic southerns                       Date: 7/29/93
Hi Yeast netters!

We have been having problems with restiction digests for southerns, and are
hoping somene will be able to help us.

We are trying to do a gene replacement in a specific strain, and to confirm the
gene replacement by southern blotting.  The problem is that our DNA
preparations (made by the method of Hoffman and Winston Gene 57:267-272 (1987))
will not cut.  This appears to be a strain specific effect, since DNA preps
from other strain backgounds cut fine.  
The DNA preps contain a white precipitate which may be polysaccharides or
carbohydrates.  Removing this precipitate by centrifugation seems to help the
digestion problem somewhat.  

Have others encountered a similar problem?  What solutions have you found for
this problem?  

Thanks.

David Stillman
Department of Cellular, Viral and Molecular Biology
University of Utah Medical Center
Salt Lake City, UT 84132
stillman at bioscience.utah.edu
phone: 801-581-5429
fax: 801-581-3607
	






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