[Histonet] Rnase free slides?

[Emily Sours]

Am I wrong in assuming that the glass slides I get (which are in a
plastic box, wrapped in plastic) are not RNase free?  They're
superfrost plus from Fisher.  It hasn't been a problem for us to
assume so, but I'm wondering what other labs think about it.
We've never treated our slides with Rnase Away or rinsed in DEPC
water.  We use DEPC water for in situ hybridization solutions.  The in
situ dishes are sprayed with RNase away, then DEPC water.
Also use OCT when you section, it'll make your life so much easier!
It's assumed in my lab that if a chemical came in and hasn't been
opened, it's RNase free.  To keep it RNase free, you always wear
gloves.  Store your slides in a new box to keep them RNase free.

RNases are everywhere on people, I assumed, and that was the problem.
I didn't think RNases are everywhere in the universe on every object.
But I could be wrong.  Anyone else have an opinion?

Emily
--
Dark Pictures, thrones and stones that pilgrims kiss
And poems that take a thousand years to die
But ape the immortality of this
Red label on a little butterfly.

-Vladimir Nabokov, concluding stanza of ‘A Discovery’ 1941.



On Wed, Jul 7, 2010 at 5:13 PM, Amos Brooks <amosbrooks <@t> gmail.com> wrote:
> Hi,
>     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 ;-)
>
> Good luck,
> Amos
>
>
> Message: 1
> 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>
>>
> Content-Type: text/plain;       charset="US-ASCII"
>
> Hello,
>
> 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.
>
> Questions:
>
> 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.
>
> Thanks!
>
> Caroline
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