Isolation of E. coli mRNA

Frank Skraly skralyf at hp-86.cae.wisc.edu
Thu Apr 13 22:31:34 EST 1995


I would like to look at the size of a few recombinant transcripts in E. coli.
There are two RNA isolation procedures that seem fairly simple in "Current 
Protocols...", one which uses as RNase inhibitors vanadyl ribonucleoside 
complex and aurintricarboxylic acid, and another that requires only DEPC.

Is inhibiting bacterial RNases really crucial (I always wind up with a ton
of RNA in minipreps if I don't add RNase), or should either of these methods
be fine?

My reason for concern is that I remember a lot of complaints about RNA
isolation and gel-running from people I thought rather highly of.

Any comments or protocols are greatly appreciated.

(Sorry if this subject has been beaten to death here, but I couldn't find
anything about it in the FAQ.)


Frank Skraly
skralyf at cae.wisc.edu



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