[Histonet] Rnase free slides?

Emily Sours talulahgosh <@t> gmail.com
Tue Jul 6 18:24:44 CDT 2010


Honestly, I think RNases are a bunch of hooha.  If you're being
careful anyway because you're doing a PCR, that should be enough.
Wear gloves, be sterile.
When I worked with a Russian post-doc, she said she did RNA in situ
hybridization without gloves and it worked.  Of course, god only knows
what her protocol was, as she had some crazy stories about Russian
labs.

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 Tue, Jul 6, 2010 at 3:49 PM, Caroline Bass <cbass <@t> wfubmc.edu> wrote:
> 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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