You can follow the method described bt Scott-Young III and used by a lot of
groups especially by Harbuz, Lightman, and our group (Garcia-Garcia,
Manzanares et al.) We use oligos (48 mer) as probes. Before ISHH we do not
do any pretreatment. After cutting the brains (12-20 µm), you do all the
treatment (including fixing with paraformaldehide).
This method serves for radiactive and non-radiactive probes. If you use
oligos as well, reply me and I will send you the protocol.
Luis (luisgarcia at altavista.net).
Dredd escribió en mensaje <352FE984.3F2093D4 at cord.ubc.ca>...
>It depends on what you are probing for - e-mail me and I can send you a
general protocol
>that we use in our labs...
>>>>gino.miele wrote:
>>> Hi, I want to do In Situ Hybridisation on mouse brain sections using
DIG-labelled
>> RNA probes. Problem is my brain tissue has been collected, frozen in
liquid N2 and
>> then stored at -80 without any pre-treatment. Can anyone give me advice
on the best
>> method to use for ISHs on brain tissue stored in this way? Thanks in
advance.
>>>> Gino Miele,
>>>>gino.miele at bbsrc.ac.uk>>>