PCR on fixed/permeabilized/sorted cells

Leigh Eward rosemarythyme80 at hotmail.com
Fri Jun 15 21:49:35 EST 2001


Karim,
     I recently came across an article entitled "Isolation of full-size mRNA
from cells sorted by flow cytometry" by C. Diez (J. Biochem Biophys Methods
1999, 40(3); 69-80) which I think could prove helpful.  They describe a
methodology in which they are able to isolate mostly intact full-size mRNA
(e.g. beta-actin) from fixed, sorted cells.
                   Leigh Eward


>From: Karim Vermaelen <Karim.Vermaelen at rug.ac.be>
>Reply-To: Karim.Vermaelen at rug.ac.be
>To: Cytometry Mailing List <cytometry at flowcyt.cyto.purdue.edu>
>Subject: PCR on fixed/permeabilized/sorted cells
>Date: Thu, 14 Jun 2001 17:42:07 +0200
>
>Hello again,
>I found only little expertise on that subject in the Cytometry archives,
>so I thought I'd raise the issue once more... Is it technically feasible
>to perform a decent RNA extraction on cells that have been fixed,
>permeabilized, labeled for an intracellular marker and FACSorted on that
>marker for several hours? Who thinks it's catastrophic, who doesn't?
>Thanks a lot for any comments/experience
>Karim
><< Karim.Vermaelen.vcf >>

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