Northern blot for low abundance mRNA

Chris LaRosa clarosa at biocomp.unl.edu
Tue Mar 7 10:24:30 EST 2000



RIGGS wrote:

> Dear readers,
>
> Does anyone have a suggestion for Northern blots for low abundance

If you cannot see a good signal using total mRNA,  then you need to prepare
fractions enriched for poly A+ mRNA using oligo dT cellulose or one of the
new matrixes for capturing poly adenylated mrna.   Lets assume that you
have a poly denylated mrna.    I have see at times that is is sometimes
necessary to use up to 20 micrograms (this is a lot) of poly A mRNA in
order to see low abundance signals.   Sometimes it is nearly impossible to
see the signal by northern blot.   More sensitive than the northern blot is
nuclease protection assays.

P. Christopher LaRosa, plant biologist





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