nuclear run-off

[David Morgenstern]

Can anybody give me any pointers on doing nuclear run-off transcription?

I'm using a protocol from Current Protocols in Molecular Biology that is 
based on the method of Greenberg and Ziff (1984 Nature 311:433).  I've also 
tried using Tri-Reagent.  In both cases I end up with very low counts in my 
RNA (<5 X 10e5).  My nuclei look good, so I don't think the problem is 
there.  Does anybody have any suggestions?  How does one figure out at which 
step things are going awry?  Thanks in advance for your help.  I can also be 
reached at Rrosenth at iupui.indycms.edu  
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