Abstract
The thymidine analogues BrdU (5-bromo-2´-deoxyuridine) and EdU (5-ethynyl-2´-deoxyuridine) are routinely used for determination of the cells synthesizing DNA in the S-phase of the cell cycle. Availability of the anti-BrdU antibody clone MoBu-1 detecting only BrdU allowed to develop a method for the sequential DNA labelling by these two thymidine analogues for determining the cell cycle kinetic parameters.
In the current step-by-step protocol, we present` two approaches optimized for in vivo study of the cell cycle and the limitations that such approaches imply: (1) determination of the cell flow rate into the G2-phase by dual EdU/BrdU DNA-labelling method and (2) determination of the outflow of DNA-labelled cells arising from the mitosis.
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Acknowledgments
The work was supported by the grant GACR 17-01897S from the Czech Science Foundation and the programs PROGRES Q26 and SVV 260371 of the Charles University, Czech Republic.
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Páral, P., Báječný, M., Savvulidi, F., Nečas, E. (2019). Cell Cycle Analysis Using In Vivo Staining of DNA-Synthesizing Cells. In: Turksen, K. (eds) Imaging and Tracking Stem Cells. Methods in Molecular Biology, vol 2150. Humana, New York, NY. https://doi.org/10.1007/7651_2019_228
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DOI: https://doi.org/10.1007/7651_2019_228
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