RE: [Histonet] Can you use RNAlater or RNAlaterice to extract RNA fromOCT-embedded tissues?

From:"Tan, MinHan"


I approached the company for an RNA protocol one year ago; the response
was that this 'application' wasn't supported.

However, I have had no problems with extraction of RNA from OCT blocks.
One can shave off multiple thick sections and then homogenize these
sections in Trizol. Alternatively, I usually trim the block by cutting
off large pieces of OCT without tissue with a razor. I subsequently
homogenize the remaining chunk of tissue mixed with OCT in Trizol. Works
great for Affymetrix microarray analysis, and there's generally no need
to slice and dice in a cryotome. :) Guess you'll just have to check to
make sure that it works for your application. 

RNA Later-ICE has worked very badly for me. The majority of tissue
samples I thawed in this product yielded degraded RNA, though I admit
that it was possible that the tissue was degraded prior to that. We had
to use it as we wanted to stick a needle into the tissue at that time. 

So just proceed with Trizol. 


-----Original Message-----
[] On Behalf Of Charles
Sent: Tuesday, November 16, 2004 2:21 PM
Subject: [Histonet] Can you use RNAlater or RNAlaterice to extract RNA
fromOCT-embedded tissues?


      I am currently working with OCT-embedded tissues and am wondering
if anyone has had success extracting RNA from OCT-embedded tissues or
has a protocol for doing so.  I am considering RNAlater-ICE by Ambion
but specifically --

(1)  is OCT fully water soluble and not infiltrative?
(2)  Should I use RNAlater-ICE as a preservative upon thawing the
OCT-embedded tissues or just proceed with TRIzol extraction?
(3)  Do I need to get rid of the OCT embedding medium and if so, how do
I do this?

    Any help would be greatly appreciated!

Dr. Charles Chiu, M.D./Ph.D.
Clinical Fellow, Infectious Diseases
UCSF School of Medicine
521 Parnassus Ave., Room C443, Box 654
San Francisco, CA  94143-0654
Cell: (415) 420-4463
Pager: (415) 719-0088
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