Density transfer as a method to analyze the progression of DNA replication forks

Methods in Molecular Biology
José Antonio Tercero

Abstract

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.

Citations

Aug 1, 2013·Nucleic Acids Research·Irene SaugarJosé Antonio Tercero
Jan 21, 2015·Scientific Reports·Esther C MorafraileMónica Segurado

Related Concepts

Carbon Isotopes
Centrifugation, Density Gradient
DNA Replication
DNA, Fungal
Nitrogen Isotopes
Genomic Hybridization
Replicon
Saccharomyces cerevisiae
Whole Genomic DNA Probes
Chromosomes, Fungal

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