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A flow cytometric method for estimating S-phase duration in plants.

Journal of experimental botany (2016-11-05)
Leigh Mickelson-Young, Emily Wear, Patrick Mulvaney, Tae-Jin Lee, Eric S Szymanski, George Allen, Linda Hanley-Bowdoin, William Thompson
RÉSUMÉ

The duration of the DNA synthesis stage (S phase) of the cell cycle is fundamental in our understanding of cell cycle kinetics, cell proliferation, and DNA replication timing programs. Most S-phase duration estimates that exist for plants are based on indirect measurements. We present a method for directly estimating S-phase duration by pulse-labeling root tips or actively dividing suspension cells with the halogenated thymidine analog 5-ethynl-2'-deoxyuridine (EdU) and analyzing the time course of replication with bivariate flow cytometry. The transition between G

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Gamborg′s B-5 Basal Medium with Minimal Organics, fine powder, suitable for plant cell culture