mRNA extractions with diatoms/celite ** mRNA? & Celite Grade? **
Dr. Duncan Clark
duncan at genesys.demon.co.uk
Fri Oct 3 03:51:26 EST 1997
In article <6125b7$17e$1 at nargun.cc.uq.edu.au>, Paul Rohde
<P.Rohde at mailbox.uq.edu.au> writes
>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!)
Having just started using the Merlin recipes, I would hazard a guess
that the small products are either binding too tightly to be eluted with
TE or water or they aren't binding at all. From memory the original
Glass Milk recipes had the same problem and it was due to non-elution.
With DNA one can take the elution component temperature up to 70C but
that might not be a good idea with RNA.
The problem with being on the cutting edge is that you occasionally get
sliced from time to time....
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