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. 2018:1703:75-86.
doi: 10.1007/978-1-4939-7459-7_5.

DNA Catenation Reveals the Dynamics of DNA Topology During Replication

Alicia Castán  1 , Pablo Hernández  1 , Dora B Krimer  1 , Jorge B Schvartzman  2
Affiliations

DNA Catenation Reveals the Dynamics of DNA Topology During Replication

Alicia Castán et al. Methods Mol Biol. 2018.

Abstract

Two-dimensional agarose gel electrophoresis is the method of choice to identify and quantify all the topological forms DNA molecules can adopt in vivo. Here we describe the materials and protocols needed to analyze catenanes, the natural outcome of DNA replication, in Saccharomyces cerevisiae. We describe the formation of pre-catenanes during replication and how inhibition of topoisomerase 2 leads to the accumulation of intertwined sister duplexes. This knowledge is essential to determine how replication forks blockage or pausing affects the dynamic of DNA topology during replication.

Keywords: 2D gels; Catenation; Replication; Saccharomyces cerevisiae; Supercoiling.

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