RE: Question RNASE

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From:"Connolly, Brett" <brett_connolly@merck.com>
To:histonet@Pathology.swmed.edu, "'Lynn Gardner'" <gardnerl@horus.ophth.uiowa.edu>
Reply-To:
Date:Fri, 09 Jul 1999 10:11:29 -0400
Content-Type:text/plain

Lynn,
Generally an extra 24 hours is not a problem if your target RNA is in high
abundance. Assuming that you have a protease step in your prehybridization
protocol you may want to include a few extra slides where your either keep
the enzyme concentration constant and extend the protease incubation time,
or increase the enzyme concentration and keep the incubation time constant.
If you prefer not to keep the tissue in fixative, wash it well in
DEPC-treated PBS.  Either way, keep at 4C.

Brett


Brett M. Connolly, Ph.D.
Senior Research Biologist
Dept. of Human Genetics
Merck Research Laboratories
WP26A-3000
PO Box 4
West Point, PA 19486
tel. 215-652-2501
FAX 215-652-2075

> ----------
> From: 	Lynn Gardner[SMTP:gardnerl@horus.ophth.uiowa.edu]
> Sent: 	Thursday, July 08, 1999 10:15 AM
> To: 	histonet@Pathology.swmed.edu
> Subject: 	Question RNASE
> 
> Hey histonetters,
> 
> Have a question about RNASE insitu we have some aorta and pulmonary artery
> tissue fixing in 4% paraformaldehyde for RNA insitu. If we have to leave
> the tissue in the paraformaldehyde over the weekend will this be a problem
> for the staining? We have left it up to 24 hours but not any longer than
> that. 
> 
> Thanks for the help.
> Lynn Gardner
> 
> 



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