Help with DD RT-PCR??

[Alex Haddad]

	HI everyone...i am at the beginning of a differential display project, 
and I was wondering if I could solicit this kind group for any helpful 
suggestions.
				I have already isolated high-quality RNA from my organism 
(Tertahymena thermophila), as assayed by Northern's and Primer 
extensions.  I am mostly concerned with the PCR steps, i.e., good primer 
combinations, appropriate controls, ect. and the post-band 
identification analysis, such as what is the most efficient and 
high-throughput way to clone DD products, confirm their induction 
patterns, ect.  I have seen many different reports in Biotechniques and 
such focusing on these topics, but I am looking for a little wider 
audience of expertise.
	I would appreciate any suggestions people might have.  Thanx in 
advance.

Alex Haddad
University of Chicago
mahaddad at midway.uchicago.edu