Abstract
DNA double-strand break (DSB) end resection is an essential step for homologous recombination. It generates 3′ single-stranded DNA needed for the loading of the strand exchange proteins and DNA damage checkpoint proteins. To study the mechanism of end resection in fission yeast, we apply a robust, quantitative and inducible assay. Resection is followed at a single per genome DSB synchronously generated by the tet-inducible I-PpoI endonuclease. An additional assay to follow resection involves recombination between two direct repeats by single-strand annealing (SSA), since SSA requires extensive resection to expose two single-strand repeats for annealing. The kinetics of resection and SSA repair are then measured using Southern blots.
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Acknowledgements
This work was supported by NIH grants GM080600 and GM125650.
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Yan, Z., Kumar, S., Ira, G. (2021). Analysis of DNA Double-Strand Break End Resection and Single-Strand Annealing in S. pombe. In: Aguilera, A., Carreira, A. (eds) Homologous Recombination. Methods in Molecular Biology, vol 2153. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0644-5_4
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DOI: https://doi.org/10.1007/978-1-0716-0644-5_4
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Publisher Name: Humana, New York, NY
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Online ISBN: 978-1-0716-0644-5
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