RE: RNA fixative

I wouldn't recommend RNAlater for any flow applications.  While it may 
preserve RNA just fine, to say that it was problematic to recover cells 
for sorting after placed in RNAlater would be a gross understatement.  It 
was sludge.

Look up my previous posts on this topic in the archive where I give gory 
details about all the things we've tried.  In the end, we just chose to 
freeze samples rather than fix at all.	While this is suboptimal for 
several reasons, it provided good flow results and sufficient RNA 
recovery.  At the time, this was the only feasible option.

There are new RNA isolation kits for retrieving from formalin-fixed 
paraffin-embedded (FFPE) tissues that have worked very well for a group 
here.  I'd be interested to see if this would work on PFA-fixed flow 
samples, but haven't had a need to try it yet.	That's where I will go 
next if this comes up again in my lab.

Dave


David McFarland
Principal Scientist
GlaxoSmithKline
----- Forwarded by David C McFarland/PharmRD/GSK on 02/21/2008 08:23 AM 
-----

"RICE,LORI P" <lrice@ufl.edu> 
19-Feb-2008 21:30
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"Cytometry Mailing List" <cytometry@flowcyt.cyto.purdue.edu>
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Subject
RNA fixative






You can try RNAlater, which was made for preserving RNA "in the 
field" at room temp.  However, for some applications, such as 
laser capture, some gene chips, some amplifications, don't work 
well with this. Check with the company to see if yours is 
compatible.


--
Lori Rice, Ph.D.
University of Florida
lrice@ufl.edu