Glass-bead RNA extraction
RBURDINE at biomed.med.yale.edu
Fri Feb 23 16:48:15 EST 1996
In <Morris_maduro-220296184953 at elegans.biology.ualberta.ca> Morris_maduro at biology.ualberta.ca writes:
> I have been trying very hard, to no avail, to get RNA from nematodes using
> the 'glass-bead' protocol found in WBG Nov '89 p.154. It is a standard
> guanidine thiocyanate/organic extraction protocol.
I have routinely used the TRIZOL reagent from Gibco-BRL to extract RNA from
worms. I have successfully used this RNA in Northen analysis, RT-PCR and RACE
TRIZOL is a reagent that contains the guanidine and the phenol. You solublize
thw worms in this solution and add chloroform to separate RNA into the
aqueous,DNA at the interface, and proteins in the organic phase. You can, in
theory, get all the components back from each of the phases but I have never
tried this. The TRIZOL and vortexing is all I have used to solublize the
worms. I did not need to use glass beads.
One advantage to using this is that the acid pH of the phenol is controlled for
by Gibco. It is basically the modified Chomczynski and Sacchi you are
currently doing. But it has worked well for me and for other people in the
lab. I'd be happy to send you a detailed protocol if you are interested.
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