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Cell Cycle Synchronization and Flow Cytometry Analysis of Mammalian Cells

  • Naoko Yoshizawa-Sugata
  • Hisao MasaiEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1170)

Abstract

Analysis of cellular DNA content and measurement of pulse-labeled newly replicated DNA by flow cytometry are useful techniques for cell cycle studies. In this chapter, we describe the protocols for cell cycle synchronization of mammalian cells, including time course designs and consideration of cell types to achieve successful experiments, along with the methods for detection of DNA. Some selected applications dealing with siRNA-mediated knockdown are also presented.

Key words

Cell cycle synchronization Flow cytometry Double-thymidine block Mitosis DNA replication Cell cycle inhibitors BrdU incorporation Click chemistry Embryonic stem cells siRNA-mediated gene knockdown 

Notes

Acknowledgments

This work was supported in part by The Naito Foundation Continuation Subsidy for Outstanding Projects (H.M.) and JSPS KAKENHI grant 23580243 (N.Y.-S.).

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.Department of Genome MedicineTokyo Metropolitan Institute of Medical ScienceSetagaya-kuJapan

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