Advice on oligo probes for non-isotopic in situ hyb

Loren Joseph, MD ljlj at midway.uchicago.edu
Fri Feb 27 13:30:31 EST 1998


I had trouble finding  appropriate msgs on this topic in the archive.
Does anyone have strong recommendations re: size of individual oligos
(and # of oligos) to use for non-isotopic in situ hybridization.  The
target  is what I hope is mod. abundance: easily detectable band on
Northern blot (32P restriction fragment probe) with overnight exposure
at -80.

Does anyone have any recommendations for the method of labelling?  Would
FITC ( to be used as a target for anti-FITC Ab) give lower background
than biotin?  How many labels per probe? 5' or 3'?

I am limited in terms of equipment and space, so telling me to just do a
plasmid prep and make an RNA probe with 35S, while possibly optimal, is
not practical advice. Any help appreciated, though.

Thank You.

Loren Joseph
University of Chicago



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