hybridization in glass tubes

suter at VAX.MPIZ-KOELN.MPG.DBP.DE suter at VAX.MPIZ-KOELN.MPG.DBP.DE
Tue Feb 18 07:46:30 EST 1992



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


###############################################################
# dr. c. suter-crazzolara    mpi for plantbreeding      koeln #
# tel --49-221-5062.221      fax --49.221.5062.213            #
# e-mail: suter at vax.mpiz-koeln.mpg.dbp.de                     #
###############################################################
 #             konstantijn had een hobbelpaard               #
  #              zonder kop en zonder staart,               #
   #              zo reed hij de wereld rond               #
    #              zo maar in zijn blote...               #
     #####################################################



More information about the Methods mailing list