[Histonet] PCR and ISH in fixed tissues

[Tony Henwood]

Caroline,

We have successfully done ISH for EBERs, Adenovirus RNA, CMV RNA,
Chromogranin mRNA, Estrogen Receptor mRNA, and nCAM RNA to name a few on
both Frozen sections and Formalin Fixed Paraffin Embedded Tissue (FFPE).

We have also successfully extracted RNA and DNA from FFPE tissues for PCR
(Ewing's and Rhabdo translocation PCR studies)and do this routinely.

Regards

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
The Children's Hospital at Westmead,
Locked Bag 4001, Westmead, 2145, AUSTRALIA.
Tel: 612 9845 3306
Fax: 612 9845 3318




-----Original Message-----
From: Caroline Bass [mailto:cbass <@t> bidmc.harvard.edu] 
Sent: Tuesday, 5 July 2005 3:54 AM
To: 'histonet <@t> lists.utsouthwestern.edu'
Subject: [Histonet] PCR and ISH in fixed tissues


Hey guys,

My boss is submitting a grant and he does not believe that you can do 
PCR (RT) or ISH with fixed tissues.  I thought you could but I don't 
know any details.  Could someone elaborate?  Is there a particular way 
the tissue should be fixed?  In other words do I have to use FFPE 
sections or can I use 4% paraformaldehyde perfusion?

Any advice would be helpful.

Thanks,

Caroline Bass


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