[Histonet] Rnase free slides?
amosbrooks <@t> gmail.com
Wed Jul 7 16:13:06 CDT 2010
This is certainly possible, but RNAase is everywhere, and in a cryostat
you really can't remove it because the RNA remover is an aqueous solution
that will freeze in the cryostat. A disposable blade (RNAase cleaned) is
probably the best you can do there. One thing to consider, I'm sure OCT
isn't RNAase free, so perhaps you should use 30% in DEPC treated water.
I have not seen RNAase free slides being sold by any vendors. I usually
use LCM slides, but your are doing this a bit differently. For this I
wouldn't waste my money on buying RNAase free slides. You can just dip the
slides in RNAase away and let them air dry (use them soon after though).
Further, I would not use charged slides as that would be counter-productive.
You are trying to get tissues off the slides, so adhesives would not be
helpful to you. Just a plain ole' glass slide with no bells or whistles.
Come to think of it that may be hard to find too ;-)
Date: Tue, 06 Jul 2010 15:49:58 -0400
From: Caroline Bass <cbass <@t> wfubmc.edu>
Subject: [Histonet] Rnase free slides?
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <C8590126.2888%cbass <@t> wfubmc.edu<C8590126.2888%25cbass <@t> wfubmc.edu>
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I'm doing RNA work for the first time. My plan is to take a fresh rat brain,
block quickly, freeze by immersing in dry-ice cooled isopentane, storing at
-80, collecting tissue sections (thickness will be determined, somewhere
between 20 and 300 um), and punching the particular regions I need out of
the sections. I will then isolate RNA from the punches for qPCR analysis.
1) does this sound like a viable plan?
2) and suggestions, what to be careful of?
3) where do I have to be careful of Rnase, should I use disposable blades,
cleaned with Rnase away?
4) where can I find Rnase free slides, or should I just make my own. I
usually use charged slides.
Any and all suggestions will be appreciated. I'm new to this and don't know
where I will have problems.
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