Help needed with in situ hybridisation

Sami Ikonen iksaka at CC.JYU.FI
Thu Jun 6 10:13:07 EST 1996


Is there anyone who has been doing in situ hybridisation with the
instruction from Wisden and Morris's "In Situ Hybridisation Protocols for
the Brain", chapter one: ISH with synthetic oligonucleotide probes ?

I tried it once, but I ran into some problems:

1) The exposure time was pretty long, 3-4 weeks. I've heard that
proteinase K -treatment would help, anyone tried that ?

2) There was pretty much non-specific background. Is that just because of
the long exposure time or should I use acetic anhydride and chloroform
treatment ?

I used uninduced and unfixed brain and 45-mer c-fos probe.

I would be very thankful for any advice,

Sami Ikonen
University of Kuopio
Finland




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