DIG-system/CSPD/high background after stripping of probe

Sibylle sohlweiler at gmx.de
Mon Feb 2 10:05:10 EST 2004


Hi!

It´s some weeks now that our DIG-detection system (Boeringer/Mannheim)
doesn´t work anymore. Our problem is the stripping of a probe before
re-hybridization it with another (stripping is absolutely essential
for our work and worked properly in the last 5 years). Although Ab-
and Substrate-Solution are always rel. new, the background is to dark
now.
The membrane which we use is HybondN (Amersham).
We checked the membrane (whose composition, besides, was changed
within the last 5 years), all solutions for hybridization were made
new, blocking-buffer was checked, time of stripping was increased.

Did "Tropix" change the composition of CSPD?

If anybody knows anything, please let us know!

Thanks in advance,
Sibylle



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