RNA recovery from cells sorted at G2 phase

Denis Snider (sniderd@fhs.csu.McMaster.CA)
Fri, 19 Jan 1996 16:22:35 -0500 (EST)

A colleague here at Mcmaster wants to sort a tumor cell line for
various cell cycle stages. In particular she wants to purify the G2
stage. In the end, RNA must be extracted from the sorted cells, so fixation
and specific DNA (not RNA) stains would be crucial.

Any suggestions on appropriate staining and fixation are welcome.
Good technical references for procedures would be welcome.
Do we need to syncronize the cells and drug treat to amplify G2
recovery?

Our setup: FACStar+, UV/488 Enterprise and HeNe lasers
We have experience in Heoscht, and PI DNA stains.

Please post all answers to the list, I'd rather not do the
summary thing.

Thanks in advance.
Denis Snider
Associate Professor
Dept. of Pathology
McMaster University

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