primers in RT in situ PCR

[Rob Wesley]

Hi.

I'm trying to work out a method for RT in situ PCR and I'm at a 
conceptual snag.  In the RT step, I want to use the 'specific downstream 
primer' but I'm not sure which one this is.  Is it the one complementary 
to the 3' end of the mRNA?  Can I use one of my PCR primers in this 
step?  Would an oligo dT primer cause me all kinds of trouble?  How long 
should the RT primer be?

Robert Wesley
Johns Hopkins University
Department of Neurology