A Proximity Ligation-Based Method for Quantitative Measurement of D-Loop Extension in S. cerevisiae
- Author(s): Piazza, A
- Koszul, R
- Heyer, WD
- et al.
Published Web Locationhttps://doi.org/10.1016/bs.mie.2017.11.024
© 2018 Elsevier Inc. 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.
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