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