mRNA ISH

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From:Huib Croes <H.Croes@mailbox.kun.nl> (by way of Marvin Hanna)
To:histonet@histosearch.com
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Dear Histonetters,

We want to perform an mRNA ISH on paraffin embedded mouse brain and
testis tissues fixed in buffered 4% paraformaldehyde solution.
So I have a couple of questions:
1. Does someone has a scheme of how to process these organs (we have a
Shandon Citadel 1000)?
2. Do we need to do the processing in RNase-free solutions (fixative,
water, alcohol) and maybe even in RNase-free xyleen and paraffin ?
3. How can we make the baskets RNase-free?
4. What is the best protocol to do an mRNA ISH and what is the best
enhancement method (tyramide??)
5. Does anybody has already did this before, if so, please email me ?

Thanks in advance

Huib Croes
Dept. Celbiology
Kath. Universiteit Nijmegen
P.O. Box 9101
6500 HB Nijmegen
email: h.croes@mailbox.kun.nl




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