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chromosome squashes for in situ hybridization

Martin Hoehne marthoe at filab.biologie.uni-freiburg.de
Thu Jun 24 10:24:22 EST 1999


Hello everybody,

i'm trying to prepare squashes from polytene chromosomes for in situ
hybridization. The freshly prepared squashes look pretty nice under the
microscope (phase contrast).

The next step in my protocol is to freeze the slides in liquid nitrogen,
flick off the coverslip, incubate the slides in Ethanol (10 min 70%,
30min 100%) and let them air dry. After this treatment all the beauty is
gone and the chromosomes look somehow shrunken. Also the banding is
hardly visible with a phase contrast microscope (a little bit better
with DIC).
Do i have to worry about this?

Thanks in advance,

Martin



Martin Hoehne
Labor Fischbach
Inst. for Biology 3
Freiburg University, Germany

marthoe at biologie.uni-freiburg.de




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