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A proximity ligation-based method for quantitative measurement of D-loop extension in S. cerevisiae
In: Methods in enzymology, vol. 601, pp. 27–44, Academic Press.
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 semi-quantitative nature and some limitations of the current PCR-based technique and facilitates the study of the recombination-associated DNA synthesis.
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