Abstract
Flow cytometry, a valuable technique that employs the principles of light scattering, light excitation, and emission of fluorochrome molecules, can be used to assess the cell cycle position of individual cells based on DNA content. After the permeabilization of cells, the DNA can be stained with a fluorescent dye. Cells which have a 2N amount of DNA can be distinguished from cells with a 4N amount of DNA, making flow cytometry a very useful tool for the analysis of cell cycle checkpoints following DNA damage. A critical feature of the cellular response to DNA damage is the ability to pause and repair the damage so that consequential mutations are not passed along to daughter generations of cells. If cells arrest prior to DNA replication, they will contain a 2N amount of DNA, whereas arrest after replication but before mitosis will result in a 4N amount of DNA. Using this technique, the role that p53 plays in cell cycle checkpoints following DNA damage can be evaluated based on changes in the profile of the G1, S, and G2/M phases of the cell cycle.
The online version of the original chapter can be found at http://dx.doi.org/10.1007/978-1-62703-236-0_20
An erratum to this chapter can be found at http://dx.doi.org/10.1007/978-1-62703-236-0_20
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Acknowledgments
The author is appreciative to Melissa Mattia-Sansorbrino and Emir Senturk for their contributions to the figures presented in this chapter. The authors are supported by grants from the National Cancer Institute (P01 CA080058, R01 CA125741, and R01 CA086001).
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Senturk, E., Manfredi, J.J. (2013). p53 and Cell Cycle Effects After DNA Damage. In: Deb, S., Deb, S. (eds) p53 Protocols. Methods in Molecular Biology, vol 962. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-236-0_4
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DOI: https://doi.org/10.1007/978-1-62703-236-0_4
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