Weird Northern again!!!

Susanne Rohrer "srohrer(removethis)" at immv.unizh.ch
Mon Jul 24 02:22:47 EST 2000



Chunxin Wang wrote:

> I used the standard hybridization condition according to the< molecular
> cloning> at 65C and washed at a high stringent condition.
>
> > > This time two new probes PCR-amplified from arabidopsis PR-1 and VSP1
> > > genes, repectively, were used and the membrane was newly transferred. I
> > > did the hybridization at the same time in different bags. However, the
> > > result was the same: all signals came from the rRNAs.

since you are working with eukaryotic RNA, have you tried isolating mRNA?

It sounds to me that the mRNA you are looking for is rare or unstable. What is
your isolation procedure - although I haven't worked with plants, there may be
sth your could improve.

What is the blotting procedure you are using? Ausubel notes that alkaline
blotting is a bad idea for rare RNA's, so we've switched to vacuum blotting
using 20xSSC.


--
Susanne Rohrer
Institute of Medical Microbiology
University of Zurich
Switzerland

srohrer at immv dot unizh dot ch







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