testis fixation/ISH/Bouins

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From:Gayle Callis <uvsgc@msu.oscs.montana.edu> (by way of histonet)
To:histonet <histonet@magicnet.net>
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To our colleague  "Down Under" and others interested in ISH technics, and
other DNA/RNA molecular biology methods (RT-PCR), Bouins is not considered
an good fixative for ISH/RT-PCR.  Acid containing fixatives cause
DNA destruction.  Formalin containing fixatives, NBF, zinc formalin, formol
saline, paraformaldehyde are fixatives of choice.  Also avoid alcohol/acetone.
Zenkers, B-5 and 2% gluteraldehyde (crosslinks too much!)
Rationale is to use crosslinking fiative to keep DNA in place.

I learned this from two ladies, Christine Bromely and Mary Maher-Sturm -
experts at doing this kind of work, in their outstanding workshop.

Gayle Callis




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