[Histonet] RE: Out of my comfort zone...

[Sue Hunter]

Hello
Just wanted to add one more thing - we actually use a dedicated pyrex dish (maybe 6x10 inches) for our water bath for RNA sections.  We use warm tap water, but you can put it in the microwave for a short bit if it needs to be warmer.  You can spray the dish with RNAse away and wipe before filling with water.
Sue

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Sarah Dysart
Sent: Tuesday, April 02, 2013 5:36 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Out of my comfort zone...

So...I have been asked to do some micro-dissection on some slides and then do downstream RT/PCR on them.  My molecular knowledge doesn't go much out of the world of IHC so...here is my question...

Has anyone ever substituted Citrate Buffer pH6 (or whatever HIER solution you are using) for proteinase K for use in RNA isolation and then later PCR?  Does this work?  The main question is will the HIER step take off the formalin linkage from the nucleic acids, or just the protein?

One last thing is what else goes into these solutions other than Citrate Buffer and Tween?  I haven't made it up in forever, I have just been ordering it from companies...I know...lazy...

Thanks

Sarah Goebel-Dysart, BA, HT(ASCP), QIHC (ASCP) Histotechnologist Mirna Therapeutics
2150 Woodward Street
Suite 100
Austin, Texas  78744
(512)901-0900 ext. 6912

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