nuclear run-on for low abundant RNA

mahayden at mahayden at
Fri Jun 28 11:14:42 EST 1996

     I'm currently setting up the nuclear runoff assay in our lab.  
You're right that it requires a lot of 32-P, but there is no getting 
around that- chemiluminescent methods don't work with this assay.  As far 
as the mess is concerned, it is no more than any other transcription or 
hybridization technique.  The protocol that I am using is one from the 
red book (Current Protocols in Molecular Biology, Unit 4.10).  The 
hardest part of the assay, by far, is the isolation of the nuclei.  If 
you are using tissue culture cells, you will need at least 5 million 
cells.  I typically use 10 million.  This assay is labor intensive 
and requires you to wait a couple of days before you know if it worked, 
but it is also a very powerful assay.  Good luck!
                                                    M. Hayden

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