hybridization in glass tubes

[suter at VAX.MPIZ-KOELN.MPG.DBP.DE]

dear netters,

somebody asked about background problems when using southern hybridization
incubators... I used these in the past, without too much problems, however
if I used glass tubes, I could get very interesting looking smears on the
exposures...a nice conversation piece, but alas not publishable.
This must have depended on the glass that I used: others have told me that
they do not have these problems at all when using glass tubes! However, I then
switched completely to plastic tubes (that is: a simple measuring cilinder for
100 ml volume and the top sawed off (most measuring cilinders have this small
extension at the top to guide the liquid)) with a cork stuck in. I could
then put in up to 20 phage containing filters at once, all on top of one
another, without any out of the ordinary background.
Since I have generally used 7% SDS in my hyb buffer, the cork may pop out if th
e
pressure goes up: then you can stick a hypodermic needle thru the cork to let
the pressure out....

Anyway, if you have background problems, it could be because of the glass and
I would advice you to do a small test with a plastic tube. This seems
especially useful, since you mentioned that you can hybridize in plastic bags
without any problems.

ciao, clemens


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