Re: RNA extraction from sorted cells

If you do choose to sort to Trizol, you need to use Trizol LS and 
precipitate using pellet paint. You'll need a least 350 ng of RNA to get 
a good pellet.

Scott W. Tighe
Senior Research Tech

Vermont Cancer Center
University of Vermont
DNA Microarray and Flow Cytometry Core Facilities
Health Science Research Facility 305
149 Beaumont Ave
Burlington Vermont 05405
802-656-2557 -DNA and Flow Lab
802-656-3936-Microarray Lab
802-656-2140 [Fax]    



Gerstein, Rachel wrote:

> 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
>
>