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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.