We often found that when we overlapped our colony/plaque lifts, even with the
mesh, they just never got washed properly. When we did not overlap, we were
always much more pleased with the background levels. If you have to overlap
because of limited space/time, we always tried to use an large volume (I know
that's not very quantitative, but we never measured, we just poured in enough to
fill about 2/3 of our hyb bottle, as opposed to using enough for about 1/4 of
the bottle when not overlapping) of wash buffer, to try and compensate, and that
seemed to help a little.
Allen Gathman wrote:
> We have been doing hybridizations for years in Tupperware sandwich
> containers, but recently bought a hybridization oven. So far, results have
> been rather disappointing. In particular, is it possible to do washes of
> lifts in the hybridization tubes? We're getting a lot of background. In
> some cases we have multiple circular lifts in a tube, stacked with some
> nylon mesh fabric separators. These in particular just don't wash very much
> at all. When we put them in sandwich containers and wash on a shaker, the
> background signal washes off as it should. Is there something we're doing
> wrong, or can you just not wash lifts in the hybridization tubes?
>> -Allen Gathman
Brian D. Cohen, Ph.D.
Laboratory of Reproductive and Metabolic Disorders
David Axelrod Institute #5031
NYS Dept. of Health
P.O. Box 22002
Albany, NY 12008-2002
cohen at wadsworth.org
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