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Date: Wed, 03 May 1995 09:34:12
From: smillshub.ansc.purdue.edu (Scott Mills)
To: angenmapiastate.edu
Subject: RNA
Message-Id: <QFA7B085millss>
Alon,
We have isolated RNA from adipose tissue for a number of years. It has
to be the worst tissue to deal with and yields are low on a tissue basis
because of all the fat. We follow the method of Chomczyinski and Sacchi
(Anal. Biochem 162:156) like most everyone else. The differences is we
incorporate a step to rid of the fat early in the procedure. Several
variations on a common theme exists. Typically, we:
4 g tiss + 10 ml Guanidinium thiocyanate (homgenize in a 30 ml
corex tube)
- centrifuge 10,000 g 4 C and carefully remove the fat cake with
a spatula. May need to briefly place the tube at -20 to harden the cake.
This is sufficient for pig fat. Rat fat is less saturated so it may not
work as well.
- proceed with remainder of the procedure
An alternative to solidification and cake removal is organic extraction
at the same step. This has been outlined by Louveau et al. (Anal. Biochem
196:308 1991).
- Once the fat is removed, the process is as for other tissues.
Good luck.
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