Help with RNA extraction method

Karen Kroeger karenmk at uwa.edu.au
Tue Aug 6 21:04:10 EST 1996


Hi, I was wondering if anyone could help - I have been having problems
extracting RNA from several different cell lines such as Hut78 and
HepG2. The RNA is degrading when I incubate at 65C in Promega's PE buffer
(primer extension kit : Tris, KCl, MgCl2, DTT, spermidine, ph 8.5).It is
not degrading in water or TE buffer, nor in PE  buffer at lower
temperatures. I've tried all sorts of things - several different pots of
PE buffer, multiple hot-phenol extractions, proteinase K digestion and
guanidinium extractions but still can't remove the problem. If anyone has
any suggestions it would be greatly appreciated. Please E-mail me at
jguy at cyllene.uwa.edu.au. Thanks heaps.

Jo Guy




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