Northern problem

[Jinglan Liu]

Hi, everyone:

I used 1.2kb cDNA probe for my Northern hyb. (Amersham easy-to-go labelling
bead). Total RNA from cultured cell, loading 30-40ug each lane. Hybridized
at 65 degree, with Clontech ExpressHyb buffer, overnight, washing at room
temp. 20 minutes twice, then 50 degree 20 minutes 1 time. Almost no
background after 72 hours exposure, and all singals went to 18S rRNA. 
What's problem? There is no repeats in my probe sequence. Do I have to use
mRNA for Northern? I really don't want to use mRNA, honestly.

By the way, I used exactly the same way to do Northern with a 5.6kb cDNA
probe, the result turned out perfect.


<http://www.biowww.net/forum/read.php?f=1&i=5336&t=5336>