[Histonet] PCR on paraffin sections

"E. Wayne Johnson 朱稳森" ewj <@t> pigsqq.org
Sun Jun 23 11:27:06 CDT 2013


We would like to start doing PCR on paraffin fixed tissues from pigs..

It has some appeal to us because we can see the lesions in the tissue
and then we can go after the causative agent post-hoc.  It may be useful
in cases where we have clear lesions on histopath but not the right 
fresh tissues
for PCR.  We are very limited on what we can do with IHC because
of the extreme difficulty of procuring the primary Ab.  It's very
simple to make PCR primers here.

We do have PCR up and running for all of the diseases that we
are interested in.  I understand that the process is basically transferring
the sections to a small tube instead of a slide, then dewax with a little
xylene, and remove the xylene with alcohol, then digest the section
to release the viral RNA.

Does anyone have any recommendations, advice, or experience with the 
step, and can suggest an appropriate enzyme mix for this digestion?

Yes I am considering in-situ hybridization but we have working PCR methods
so that seems to be a simple logical step.

E. Wayne Johnson
Enable Ag Tech

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