polytene in situs

John Emery jemery at mail.sas.upenn.edu
Mon Jan 30 18:59:28 EST 1995


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!



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