Help with RNA extraction method

Ned Mantei mantei at
Wed Aug 7 02:01:59 EST 1996

In article <4u8tiq$gvg at>, karenmk at (Karen
Kroeger) wrote:
 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).

Probably alkaline hydrolysis--pH 8.5 is already alkaline enough to be
unhealthy for RNA over the long term, and incubation at 65 C makes the
reaction go much faster. Cleaning up the RNA would in this case not help.

Ned Mantei
Dept. of Neurobiology, Swiss Federal Institute of Technology
CH-8093 Zurich, Switzerland
mantei at   Fax: +41-1-633-1046

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