fixation for RT-PCR
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From: | Gayle Callis <uvsgc@msu.oscs.montana.edu> (by way of histonet) |
To: | histonet <histonet@magicnet.net> |
Reply-To: | |
Content-Type: | text/plain; charset="us-ascii" |
My SLC NSH convention workshop recommended
NBF, 10% aq
4% paraformaldehyde
10% NBF
formal saline
ZN formalin works for ISH.
Fix fresh tissues 8 hours to 3 days.
Rationale: Cross linking fixatives keep DNA in place
Avoid: Bouins, 2% gluteraldehyde, Zenker, B-5, acetone/alcohol,
Rationale: any heavy metal or picric acid destroys DNA., Glut is a
crosslinker,
but the PCR reagents cannot penetrate.
Great workshop by two gals who REALLY knew their RT-PCR!
Gayle Callis
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