Re: In-situ microtomy

From:"J. A. Kiernan" <jkiernan@julian.uwo.ca>

On Wed, 6 Dec 2000, Patrick M. Haley  wrote:

> I have  a researcher wanting to perform in-situ on mouse feet.
> The specimens will be fixed in Methyl-Carnoy's. I have two questions:
> 
> Can these be decalcified without loosing DNA/RNA, and what is 
> the reagent of choice?

DNA should be OK. RNA can be hydrolyzed and extracted by 
exposure to acids. I think messenger RNA would be more
easily removed than ribosomal RNA. The correct decalcifying
agent for your job would be disodium EDTA at pH about 7.5.

 John A. Kiernan,
 Department of Anatomy & Cell Biology,
 The University of Western Ontario,
 LONDON,  Canada  N6A 5C1





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