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A Live-Cell Imaging Approach for Measuring DNA Replication Rates.

Cell Rep. 2018;

Dovrat D, Dahan D, Sherman S, Tsirkas I, Elia N, Aharoni A.

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Recombinant Proteins	Unless otherwise stated, all experiments were conducted on populations that were synchronized in G1 phase by addition of 10 mg/mL of a-factor (GenScript) for 2 hr.	Get A Quote

摘要

We describe a simple and direct approach to measure the progression of single DNA replication forks in living cells by monitoring two fluorescently labeled loci downstream of an origin of replication. We employ this approach to investigate the roles of several leading and lagging strand factors in overall replisome function and show that fork progression is strongly dependent on proper maturation of Okazaki fragments. We also demonstrate how related cellular phenotypes, such as cell-cycle progression and the dynamics of sister chromatid cohesion, may be simultaneously monitored and correlated to DNA replication at the single-cell level.,Copyright ? 2018 The Author(s). Published by Elsevier Inc. All rights rese... More