GIT RNA prep from bacteria.

[Brad Nicholson]

In article <32519D66.7BD7 at ss.gu.se>, John-Olov Jansson <joj at ss.gu.se> wrote:

> Hello,
> 
> I have one question regarding the Chomczynski-Sacchi method for RNA prep
> with GIT: Do you need to modify the protocoll for bacteria? 
>snip<

Hi John-Olov;

It is the experience of several people in our lab that to get reliable RNA
from E. coli or Salmonella you need to use a hot phenol protocol.  It
seems that the average mRNA half-life is just to short or the commercial
methods don't recover all of the transcripts quantitatively.  I personally
didn't have luck with the C-S protocol and another person in the lab
didn't have satisfactory results with C-S or the Qiagen RNA isolation
kit.  It's a pain, but I can isolate 1.2-1.5 mg of RNA that is in good
shape.

If you are interested let me know and I'll post a copy.

Best of luck,
Brad
=====================================================================
Brad Nicholson                      |"If it worked the first time, 
Department of Pathology             | it wouldn't be research.'
University of Utah                  | ...Brad Nicholson
Salt Lake City, UT 84132            | My opinions are solely my own.
brad_nicholson at hlthsci.med.utah.edu |
or: (801)-581-4365                  | iligitimi non corborundrum
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