mRNA in situ hybridization

[littgj at esvx17.es.dupont.com]

In article <4125u7$ri9 at sifon.cc.mcgill.ca>, jim at parasit.lan.mcgill.ca writes:
>I want to do mRNA in situ hybridization using dna oligonucleotide probes to nematode sections fixed and 
>embeded in wax. Does anyone have any experience with this type of in situ hybridization and know the 
>relative senistivity of this type of assay? Any help would be appreciate
We have worked out an amplification system
that has been applied to the nonradiometric 
detection of RNA (See Raap et al, Ultrasen-
sitive FISH using peroxidase mediated deposition
of biotin or fluorochrom tyramides, Human
Molecular Genetics, 4 (1995), pp529-534). In 
our lab, using probes to muscarinic receptor
mRNAs in the hippocampus of rat brain tissue 
sections, the sensitivity of either the biotin
format with DAB chromogenic detection or the
direct fluor deposition was at least as good
in less than a day of work as compared to the
need to use radioautography with a sulfur-35 
probe for a month.  Regarding the issue of 
paraffin embedded samples, you might also want
to check Kertstens et al, A Novel In Situ 
Hybridization Signal Amplification Method based
on the Deposition of Biotinylated Tyramine,
J. Histochem. Cytochem 43, 347-352 (1995).
 
Contact me if you have any interest. 



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