mRNA extractions with diatoms/celite ** mRNA? & Celite Grade? **

Paul Rohde P.Rohde at mailbox.uq.edu.au
Fri Oct 3 02:03:03 EST 1997


Hi,

I'm trialling the the N.A. extraction method of Boom et al [1] based
on diatamaceous earth/celite

Is this method Ok for mRNA extractions?

On a dummy trial, extracting total RNA as starting material (obtained
from cell lines with TRI reagent) the method yeilds rRNA beatifully,
but background mRNA smaller than the rRNA bands are much decreased,
and gets worse with smaller RNA until there is no tRNA at all.
(The starting total RNA has tRNA as the brightest part on a gel!)

I will be using a lot of celite for each extraction (on ejaculate) so
I opted to buy the much cheaper 95% Celite(R) 521 (Aldrich) (muddy
looking colour) instead of the white analytical grade.  ** Is this a
problem or is celite not good for mRNA extraction? **  Any advances on
Booms et al method for the laboratory?



[1]  Boom et al "Rapid and Simple Method for Purification of Nucleic
     Acids"  J. Clin. Micro. (1990) 28(3) p495.



Thanks in advance

Paul R. Rohde
Dep of Surgery, Univ. Qld., Royal Brisbane Hosp. Australia.
aus.net at usa.net




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