[Histonet] RE: Paraffin sections for molecular assays

[Helen Fedor]

Hello, For DNA, we use a new blande and we wipe down microtome, handles, front of blade holder with 95% ethanol on a gauze sponge. For RNA,  use a new blade and wipe down all with RNAzap including the water bath and rinse with DEPC water. Use  clean water in bath, distilled will work or DEPC water. Depending on the stringency of the assay. You can sometimes get away with a little less. But some of the assays that are being run can cost up to $1,000.00. So the time spent cleaning up is justified. Make sure you are wearing gloves for both RNA and DNA, during the whole process, even when handling the slides .(including the process of labeling slides). I usually lay the slides out on pape rtowels on the lab bench while labeling them.

Helen Fedor
Johns Hopkins University
TMA and Prostate Spore Lab Manager
JHU SOM Uro/Pathology
600 N. Wolfe Street, Marburg Room 406
Baltimore MD, 21287-7065
410-614-1660 / Fax 410-614-3695
Pager 410-283-3419


________________________________________
From: histonet-bounces <@t> lists.utsouthwestern.edu [histonet-bounces <@t> lists.utsouthwestern.edu] on behalf of Crowell, Thomas [thomas.crowell <@t> novartis.com]
Sent: Tuesday, October 04, 2011 12:44 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Paraffin sections for molecular assays

Dear Histonetters,

Can you tell me what procedures you use to prevent DNA/RNA contamination of tissue sections when handling multiple samples.  Do you just wipe down blade holder and blade (or use a new blade) between samples, or do you use more stringent cleaning?


Thomas Crowell
Diagnostic Development
Novartis Molecular Diagnostics
45 Sidney Street
Cambridge, MA 02139
USA

Phone    +1  617 8717460
Fax         +1  N.A.
thomas.crowell <@t> novartis.com<mailto:thomas.crowell <@t> novartis.com>
www.novartis.com<http://www.novartis.com/>


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