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

[Mark Tarango]

We use a heat block during digestion.  There is less chance of
contamination with a heat block than with a water bath.

... and no we don't use antigen retrieval solution for this!  We use a
Qiagen kit too.

Mark


On Wed, Apr 3, 2013 at 9:11 AM, Helen Fedor <hfedor <@t> jhmi.edu> wrote:

> We have been using store bought gallons of distilled water in our water
> baths. This water has been boiled so enzyme activity should be absent.
>
>
> Helen
>
> 410.614.1660
>
> http://tmalab.jhmi.edu/
> http://prostatebiorepository.org/
>
>
>
> Helen
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:
> histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Sue Hunter
> Sent: Wednesday, April 03, 2013 10:55 AM
> To: Sarah Dysart; histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] RE: Out of my comfort zone...
>
> 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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