RE: <Suspected Spam> RNA extraction from sorted cells

We sort directly into Trizol and just follow their protocol.   You should be able to use Trizol to isolate RNA from cells that were frozen, it will just depend on whether there was degradation of the RNA, ie if they were snap-frozen in a dry ice/ETOH bath and stored at -80, this should work IF you sorted enough cells.

e-mail me off the list if you have questions

=======================================================
Rachel M. Gerstein, Ph.D.
Associate Professor
Department of Molecular Genetics and Microbiology
Graduate Program in Immunology/Virology
University of Massachusetts Medical School
55 Lake Avenue North
Worcester, MA 01655-0002
(508) 856-1044
(508) 856-5920 (FAX) 



-----Original Message-----
From: Julie Bertout [mailto:julie.bertout@ibl.fr]
Sent: Thu 3/27/2008 8:30 AM
To: cyto-inbox
Subject: <Suspected Spam> RNA extraction from sorted cells
 
Hello,
We sorted splenic cells to have different subsets of lymphocytes, 
pelleted cells and froze them. We tried to extract RNA (using a Qiagen 
kit) from these but had no results...

After further investigation, I found out that the best to have a good 
RNA extraction from sorted cells was to sort cells directly in the lysis 
buffer (I found few posts about this on the purdue list). We will do it 
next time (if you have protocols or products that works fine, please 
tell me).

But is there any way to extract RNA from the cells we sorted? Does 
someone have a protocol adapted to sorted lymphocytes which have been 
frozen?

Thanks a lot,

Julie Bertout
Flow cytometry Core
Institut Pasteur de Lille - Institut de Biologie de Lille
France