in situ hybridization
tobin at ncifcrf.gov
Thu Mar 21 13:24:43 EST 1996
Our lab is getting into in situ hybridization to identify tissues containing
specific DNA or RNA. This would entail standard in situ hybridization,
PCR in situ, and RT-PCR in situ. There are many, many protocols out there
for doing these experiments. At first, I would like to try in situ hyb with
a radioactive probe and then move to a fluorescent probe. The PCR-based assays
would be fluorescent.
Can I get some opinions on how to make the best probe? How long should it be
for in situ hyb? Would a random primed or PCR-generated probe have higher
sp. activity than the nick-translation probes? If the target is 2.5 kb,
would it be better to use a 2.5 kb probe or an 0.5 kb probe (the smaller probe
might penetrate and bind better).
Any advice would be appreciated.
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