A Proximity Ligation-Based Method for Quantitative Measurement of D-Loop Extension in S. cerevisiae

Aurele Piazza (1, 2) , Romain Koszul (1) , Wolf-Dietrich Heyer (2)

1 Régulation spatiale des Génomes - Spatial Regulation of Genomes
2 UC - University of California

Homologous recombination faithfully restores the sequence information interrupted by a DNA double-strand break by referencing an intact DNA molecule as a template for repair DNA synthesis. DNA synthesis is primed from 3'-OH end of the invading DNA strand in the displacement loop (D-loop). Here, we describe a simple and quantitative proximity ligation-based assay to study the initiation of homologous recombination-associated DNA synthesis initiated at the D-loop and final product formation. The D-loop extension assay overcomes the semiquantitative nature and some limitations of the current PCR-based technique and facilitates the study of the recombination-associated DNA synthesis.

Keywords: D-loop; DNA synthesis; DSB repair; Homologous recombination

Domains: Life Sciences [q-bio]

Aurele Piazza, Romain Koszul, Wolf-Dietrich Heyer. A Proximity Ligation-Based Method for Quantitative Measurement of D-Loop Extension in S. cerevisiae. Mechanisms of DNA Recombination and Genome Rearrangements: Intersection between Homologous Recombination, DNA Replication and DNA Repair, 601, pp.27-44, 2018, Methods in enzymology, 978-0-12-813979-0. ⟨10.1016/bs.mie.2017.11.024⟩. ⟨pasteur-02868112⟩