freezer storage of lymphocytes (buffy coats) prior to DNA extraction

Richard P. Grant rpg14 at yahoo.co.uk.invalid
Tue Jan 22 07:38:54 EST 2002


In article <%KY28.8234$E%2.1371252839 at newssvr16.news.prodigy.com>,
 "bprman" <bprman at hotmail.com> wrote:

> "I note that there is a misspelling in my earlier message regarding freezing
> of cells before DNA extraction. The question was whether it would be OK to
> freeze the cells at minus 80 degrees centigrade immediately after plasma is
> extracted without freezing medium, which uses up a lot of FCS. Please
> comment."
> 
> I seem to recal that this is fine, but since we have always worked with
> whole blood, I need help answering the question.

Your collaborator should be fine freezing buffies without FCS.  I know a 
genomic DNA extraction service that has successfully extracted from 
frozen buffies (the vampire slayers??).  Indeed, the first stage of the 
traditional salting-out protocol is essentially a buffy prep.

Of course, they could always do a pilot experiment . . .


hth,

r

-- 
Richard P. Grant  0x5F9559B1                         MRC Lab of Mol Biol
rpg 'at' mrc-lmb.cam.ac.uk   http://www2.mrc-lmb.cam.ac.uk/personal/rpg/
   'When women are depressed they either eat or go shopping.
         Men invade another country' ‹ Elayne Boosler




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