RNA Slot-blotting woes
ez008413 at dale.ucdavis.edu
Tue Jun 4 17:45:14 EST 1996
I'm trying to quantify my gene's transcript in several tissues using
slot-blotting of total RNA and known amounts of target DNA as standards.
My previous northerns show comparable high level expression let's say in
tissues a,b and c. When I slot-blotted the same amount of total RNA from the 3
tissues as I ran for the northrens, I got starkly different results:
such as, only tissue a showed signal with b and c showing nothing. In short,
the northern data and slot-blot data are inconsistent. Because my northerns
have been reproducible, I suspect something is wrong with the
slot-blotting. So, in my subsequent attempts, I've turned down the vacuum
and also added carrier tRNA to improve RNA retention on
the blot. Still no improvement. Could somebody help me troubleshoot? I'd
appreciate any advice.
More information about the Methods