Reply to "Total RNA from fibroblasts"

nishir at ohsu.edu nishir at ohsu.edu
Fri May 28 17:34:57 EST 1993


In article <1993May28.085739.7245 at gserv1.dl.ac.uk>
GRGGTA at PATERSON.CHRISTIE-HOSPITAL.MANCHESTER.AC.UK writes:
>Dear all,
>         The method in use in our lab to prepare total RNA from fibroblasts
>and lymphocytes is extremely reliable and easy (though inevitably not that
>inexpensive at 200 pounds sterling per 200ml - 10^7 fibroblasts can be
processed with 2-3ml ie 2-3 pounds/$5 per isolation). We use a product sold by
Biogenesis Ltd of 12 Yeomans Park, Yeomans Way, Bournemouth, England BH8 OBJ,
UK, called 
>RNAzol. This is simply a guanidinium thiocyanate extraction which has been
>pre-prepared as a single solution which is added directly to the cell
monolayer
>or pellet. This lysate is gathered into a 15ml polypropylene tube, 0.1 volumes
>of chloroform added and mixed well. After 15min this is spun at 12kxg and the
>upper (colourless) aqueous layer pipetted into a clean tube. An equal volume
>of isopropanol precipitates the RNA. This is pelleted at 8kxg, washed once
with 75% ethanol, dried and is then ready to use. Hope this is some use. I also
hope
>this gets on to the bulletin board as I am a new "netter" with no previous 
>experience!!!
>
>FAX number for Biogenesis is +44 202 530367 (international)
>                             (0202) 530367
>               Telephone     +44 202 522895 (international)
>               Telephone     +44 202 522895 (international)
>                             (0202) 522895 (UK)
>
>NB I am not a sales rep for this company-though I have "sold" it to several
>departments in this institute!
>
>Graham Atherton
>

For American subscribers, you might be interested to know that the same
solution (acid guanidinium extraction) is sold by Molecular Research Center,
Inc. 5645 Montgomery Rd., Cincinnati, OH 45212. 1-800-9868; FAX 513-841-0080. 
The product is called TriReagent and the protocol for RNA extraction is
essentially the same.  We get very good yields of RNA with it: 4-5- fold better
than other techniques.  Whenever we've called the company for help they have
been very helpful and we have always talked with P. Chomczynski who was an
author of this technique when it was first published.  I assume he owns the
company. (I do not work for the company).

Rae Nishi
OHSU
Portland, OR



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