Problems with RNA In-situ Hybridization

Frank Gerlach frankgerlach245 at
Sat Sep 1 03:12:50 EST 2001

Dear newsgroup,

I have some problems regarding to RNA-RNA in-situ hybridisations.
I get no specific labeling with my antisense probe, produced by the DIG-RNA
labeling Kit from Roche. The hybridization temperature is 50 °C. We tried
the probes at paraffin-embedded tissues treated with proteinase K and also
at tissues that were cut with a cryostat.
I tried several protocols but it would be nice if someone could send me his
or her protocols.
I am not sure if our sense and antisense probes work properly. To  test them
I have put them on a formaldehyd-gel. One probe shows one distinct band. The
other one also has a band at the same size but there is smear around it.
Have you any experience which "gel-picture" of the probes is better?

Thanks a lot in advance.

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