A Proximity Ligation-Based Method for Quantitative Measurement of D-Loop Extension in S. cerevisiae.
- Author(s): Piazza, Aurèle
- Koszul, Romain
- Heyer, Wolf-Dietrich
- et al.
Published Web Locationhttps://doi.org/10.1016/bs.mie.2017.11.024
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.