Riboprobes, Northerns, and high background

Karl Fischer tyr-2 at bones.biochem.ualberta.ca
Wed Nov 20 12:20:25 EST 1996

In article <56v7mj$23ic at r02n01.cac.psu.edu>, kgilbert at cmp.hmc.psu.edu
(Gil) wrote:

> I detect single bands on Northerns, but get high lane background.  Any
> suggestions on ways to eliminate this high backgorund would be greatly
> appreciated.  Thanks in advance.

One thing we've found (and other have posted similarly) that the blocking
components of your hyb solution can change depending on the type/brand of
membrane you use. One such membrane distributer suggests increasing SDS to
1 % final while another retains SDS at 0.1% (standard for our Southerns)
yet increases Denhardt's solution to 10X final concentration rather than

Bottom line - take a look at the distributer's recommendations for the
membrane (a la Schleicher and Schuell's Product Guide and Methods - no
affiliation). If there's nothing in the catalogue or accompanying the
product call their tech rep and they will usually fax you the info on the
same day.


Karl the hepB guy

Karl Fischer
tyr-2 at bones.biochem.ualberta.ca

More information about the Methods mailing list