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HELP! Bacterial mRNA isolation

Emir Khatipov ekhatipov at NOuchicago.edu
Mon Mar 15 12:49:59 EST 1999


I would appreciate any guides on the following:


1) How to rapidly quench BACTERIAL cells for subsequent RNA isolation?
Convential cell harvesting by centrifugation is too long. Would treatment
with trichloracetic or chloric acid work? Alkaline treatment?


2) How to separate BACTERIAL mRNA from total RNA (mRNA+rRNA)? I.e. how to
get rig of rRNA?
Bacteria DO NOT HAVE poly-A tails, thus commercial kits based on
poly-A-matrix binding do not work. In fact, there are poly-A in bacteria,
but not all mRNA have them.


I need mRNA for expression analyses.


Thank you.


Emir








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