QuickHyb (Stratagene): Northern blot problems????

tsg1 at york.ac.uk tsg1 at york.ac.uk
Tue Nov 18 08:55:49 EST 1997


Hi,

I have used Stratagens'e QuickHyb buffer for northerns for awhile with no
problems. However, recently
(since using a new bottle of QuickHyb) i have experienced high backgrounds
on the northerns. It appears to
be related to the QuickHyb blocking/hybridization step, since I usually
mark the position of the RNA makers
with a pen - and these marks are now clearly visible (white) on a grey
background. I use BioMax MS (high
sensitivity film) with o/n exposure and get grey blots. I've used Quickhyb
with BioMax exposure
times of 5 days with no problems before (a blot done only a week ago).
Since I'm looking for a relatively rare
message- the high background means i cannot detect it!!! Using a probe to
a highly expressed gene is able
to detect message - but the background is still visible!!!

I was wondering if anyone has experienced this type of problem before and
if they have any suggestions
to reduce or overcome them? 
Anyone had experience with other company's rapid hybridization buffers
and have good/bad comments??

Your comments welcome and thanx for your time.

email:
tsg1 at york.ac.uk

-- 
T. S. Grewal
Bone & Joint Research Group
Department of Biology,
University of York,
York,  UK.

email: tsg1 at york.ac.uk



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