Cell Synchronization by Double Thymidine Block

Cell synchronization is widely used in studying mechanisms involves in regulation of cell cycle progression. Through synchronization, cells at distinct cell cycle stage could be obtained. Thymidine is a DNA synthesis inhibitor that can arrest cell at G1/S boundary, prior to DNA replication. Here, we present the protocol to synchronize cells at G1/S boundary by using double thymidine block. After release into normal medium, cell population at distinct cell cycle phase could be collected at different time points.

Keywords: Cell cycle; Cell synchronization; DNA replication; DNA synthesis; Thymidine.

Conflict of interest statement

Competing interests The authors have declared that no conflict of interest exists.

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References

    1. Chen G., Magis A. T., Xu K., Park D., Yu D. S., Owonikoko T. K., Sica G. L., Satola S. W., Ramalingam S. S., Curran W. J., Doetsch P. W. and Deng X.(2018). Targeting Mcl-1 enhances DNA replication stress sensitivity to cancer therapy. J Clin Invest 128(1): 500-516. - PMC - PubMed
    1. Graña X. and Reddy E. P.(1995). Cell cycle control in mammalian cells: role of cyclins, cyclin dependent kinases(CDKs), growth suppressor genes and cyclin-dependent kinase inhibitors(CKIs). Oncogene 11(2): 211-219. - PubMed
    1. Ho Y. S., Duh J. S., Jeng J. H., Wang Y. J., Liang Y. C., Lin C. H., Tseng C. J., Yu C. F., Chen R. J. and Lin J. K.(2001). Griseofulvin potentiates antitumorigenesis effects of nocodazole through induction of apoptosis and G2/M cell cycle arrest in human colorectal cancer cells. Int J Cancer 91(3): 393-401. - PubMed
    1. Koç A., Wheeler L. J., Mathews C. K. and Merrill G. F.(2004). Hydroxyurea arrests DNA replication by a mechanism that preserves basal dNTP pools. J Biol Chem 279(1): 223-230. - PubMed
    1. Léger K., Hopp A. K., Fey M. and Hottiger M. O.(2016). ARTD1 regulates cyclin E expression and consequently cell-cycle re-entry and G1/S progression in T24 bladder carcinoma cells. Cell Cycle 15(15): 2042-2052. - PMC - PubMed