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genomic southerns

odile%FRCGM51.earn at uk.ac.earn-relay odile%FRCGM51.earn at uk.ac.earn-relay
Fri Aug 6 13:45:53 EST 1993

Hello David and yeast netters !
David Stillman wrote a long time ago :-)..
>We have been having problems with restiction digests for southerns, and are
>hoping some one will be able to help us.
> 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.
For yeast genomic DNA extraction, we use the technique described by in
"methods in yeast genetics" by Rose, Winston and Hieter Cold spring
harbour laboratory (1990) p 128- 129.
This method consists to do spheroplasts by zymolyase,
DNa precipitation, RNAse treatement and no phenol extraction. In our hands
this is the best method for genomic DNA extraction and we have no problems
for enzymatic digestions.
I hope that it could help you, this a nice method
(David, if you do not find the CSH manual, let me know, I shall send you
the protocole by fax .... ;-))
Odile Ozier-Kalogeropoulos
91190 Gif sur Yvette

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