Reply to "Total RNA from fibroblasts"

GRGGTA at PATERSON.CHRISTIE-HOSPITAL.MANCHESTER.AC.UK GRGGTA at PATERSON.CHRISTIE-HOSPITAL.MANCHESTER.AC.UK
Fri May 28 04:53:00 EST 1993


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



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