Summary
The density transfer technique is a valuable tool to examine the progression of individual DNA replication forks. It is based on the transfer of cells from a medium containing dense isotopes to a medium with light (normal) isotopes (or vice versa), to obtain DNA sequences hybrid in density that can be identified as replicated molecules. Using specific DNA probes along a chromosome, the dense isotope transfer method allows determining the extent of replication at any position of a replicon and the rate of replication fork progression. In the eukaryotic model budding yeast, this technique has been useful to establish a role for different proteins during the elongation of chromosomal replication and to analyze the movement and stability of DNA replication forks under different experimental conditions.
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References
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Acknowledgments
The author would like to thank John F.X. Diffley for his encouragement and help in developing this method to study DNA replication fork progression, and Kristine Bousset for her introduction to the density transfer technique. J.A.Tercero’s work is funded by grants BFU2007-67445 and Consolider CSD2007-00015 from the Spanish Ministry of Education and Science (MEC), and by an institutional grant from the Fundación Ramón Areces to the Centro de Biología Molecular Severo Ochoa.
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Tercero, J. (2009). Density Transfer as a Method to Analyze the Progression of DNA Replication Forks. In: Vengrova, S., Dalgaard, J. (eds) DNA Replication. Methods in Molecular Biology, vol 521. Humana Press. https://doi.org/10.1007/978-1-60327-815-7_11
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DOI: https://doi.org/10.1007/978-1-60327-815-7_11
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