Isolation of mammalian nuclei for nuclear run-off assay - (Apr/16/2002 )

Hi...

I'm trying to isolate nuclei from HeLa cells for nuclear run-off assays.  Using my current protocol, the nuclei would lyse (becomes extremely viscous) when I add the transcription buffer for the assay.   Does anyone have a successful protocol or tips?

Add RNase Free DNAaseI. That will take care of your viscous DNA and you will get a decent radilabelled RNA.

Ranjit K. Giri.
ranjitgiri@hotmail.com