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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.
Abstract

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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