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:

<snip> 
> 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
5X.

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.

Cheers

Karl the hepB guy

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





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