Northerns...AGAIN!

Deb Britt Deborah_Britt at brown.edu
Wed Oct 8 14:55:32 EST 1997


Lots of things can go wrong with Northerns, but the first question I would
ask is: is the problem with the probe, or something with the technique. 
You need to get ahold of a plasmid that has a "housekeeping" gene, 
expressed in all cells in your population    (one of the labs here uses
GAPDH).  Label and hyb that, and if nothing shows up then there is a
problem with your RNA or blotting/hyb technique.  If that works, start
looking into what might be wrong with your probe.  Hybridization with a
housekeeping gene probe is routinely used anyway, to demonstrate that all
wells contain the same amount of RNA, and that differences you see with
your probe of interest really are differences in expression.

Good luck

Deb Britt

-- 
Deborah Britt, Ph.D.
Department of Medical Oncology
Rhode Island Hospital




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