[Histonet] Pancreas RNA

[Jacqui Detmar]

Hey there.  I *personally* haven't done any RNA work for LCM on
pancreatic tissue, but I have listened to two (independent) researchers
- who work on mouse pancreas - rave about how RNAlater (Ambion) saved
their sanity.  Another researcher really liked the LCM staining kit
(also from Ambion) b/c it prevented the activation of endogenous
proteases through the use of non-aqueous solutions.  Again, I haven't
used these techniques, but I've heard how difficult the pancreas can be
to deal with and I thought I would pass these on as a possibility.

Good luck,

Jacqui


Jacqui Detmar, Post-doctoral Fellow
Samuel Lunenfeld Research Institute, room 876
Mount Sinai Hospital
600 University Avenue
Toronto, ON, Canada
M5G 1X5

phone:   416-586-4800 x2451/x2290
fax:        416-586-8588
email:     detmar <@t> mshri.on.ca


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
Madeline.Branflick <@t> sanofi-aventis.com
Sent: Thursday, June 05, 2008 3:12 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Pancreas RNA

Hello:
Does anyone have experience working with pancreas tissue for downstream
LCM?  Particularly FFPE and frozen collection procedures/protocols.  We
have tried both formats, and were unsuccessful in maintaining RNA
integrity.  We've tried to get around the endogenous ribonuclease issue
by snap freezing in chilled isopentane, or a direct fixation in 10% NBF.
Any help is greatly appreciated.
Madeline
 
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