RE: In Situ with Bone

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From:"Connolly, Brett" <> (by way of histonet)
Date:Wed, 20 Oct 1999 03:46:51 -0400
Content-Type:text/plain; charset="us-ascii"

You'll probably have to bite the bullet on this one. mRNA, being the labile
rascal that it is, would most likely not survive any acid decalcification.
If you are unsure of the abundance of your target, life gets even more
difficult. How will you know if there is any of your target mRNA left after
acid treatment?  Maybe some of the 'boneheads' can recommend an embedding
medium as you alluded to. If you go plastic, of course, your going to have
to deal with getting your probe in.
I'm a big fan of tyramide signal amplification so what ever route you go, if
you think you may have destroyed some of the target mRNA or there is very
little there to begin with, I would recommend TSA (get out your wallet).


Brett M. Connolly, PhD.
Senior Research Biologist
Dept. of Pharmacology
Merck Research Laboratories
PO Box 4
West Point, PA 19486
tel. 215-652-2501
FAX 215-652-2075

> ----------
> From: 	Jay Turner[]
> Sent: 	Monday, October 18, 1999 1:06 PM
> To:
> Subject: 	In Situ with Bone
> Does any one have any suggestions for a time-efficient in situ
> hybridization protocol for bone specimens?  I was hoping to avoid
> the long, painful process of EDTA decalcification, if possible.
> Would a dilute formic acid decal solution be gentle enough for mRNA
> preservation?  Is there an embedding medium for nondecalcified bone
> that is suitable for in situ?  Any help would be greatly
> appreciated.
> Sincerely,
> Jay D. Turner
> Dept. of Orthopaedics
> University of Medicine and Dentistry of NJ
> Newark, NJ
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