From owner-drosophila@net.bio.net Mon Jan 30 22:00:00 1995
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From: jemery@mail.sas.upenn.edu (John Emery)
Newsgroups: bionet.drosophila
Subject: polytene in situs
Followup-To: bionet.drosophila
Date: 30 Jan 1995 23:59:28 GMT
Organization: University of Pennsylvania
Lines: 13
Message-ID: <jemery-300195195353@guildlab.sas.upenn.edu>
NNTP-Posting-Host: guildlab.sas.upenn.edu

I'm looking for help with in situ hybridization of cDNAs to polytene
chromosomes. I have had reasonable success using a 3 kb cDNA, but a 0.5 kb
subclone of this will not work. 

Is it possible to do in situs with cDNAs < 0.5 kb?  Any suggestions?

Here are some specifics of the protocol I'm following. Probes are uncut
plasmids nick translated with bio-16-dUTP. Hybridization is in a 50%
formamide buffer at 37C overnight. Washing is done at 53C. I'm using a
1:250 dilution of streptavidin-HRP complex (ENZO Detek-1-HRP kit) and DAB
for detection.

Thanks in advance!

.