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[Protocol] CC-seq: nucleotide-resolution mapping of Spo11 DNA double-strand breaks in S. cerevisiae cells

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journal contribution
posted on 2024-09-02, 10:39 authored by George BrownGeorge Brown, William GittensWilliam Gittens, Rachal M Allison, Antony OliverAntony Oliver, Matt NealeMatt Neale
During meiosis, Spo11 generates DNA double-strand breaks to induce recombination, becoming covalently attached to the 5' ends on both sides of the break during this process. Such Spo11 "covalent complexes" are transient in wild-type cells, but accumulate in nuclease mutants unable to initiate repair. The CC-seq method presented here details how to map the location of these Spo11 complexes genome-wide with strand-specific nucleotide-resolution accuracy in synchronized Saccharomyces cerevisiae meiotic cells.<p></p>

Funding

Elucidating Eukaryotic Topoisomerase III Activity: An Enzymatic Double-edged Sword : BBSRC-BIOTECHNOLOGY & BIOLOGICAL SCIENCES RESEARCH COUNCIL | BBSRC – Grant R

Elucidating Eukaryotic Topoisomerase III Activity: An Enzymatic Double-edged Sword : Biotechnology and Biological Sciences Research Council | BB/V005081/1

Spatiotemporal control of meiotic recombination : WELLCOME TRUST | 225852/Z/22/Z

Spatial regulation of meiotic recombination : WELLCOME TRUST | 200843/Z/16/Z

History

Publication status

  • Published

File Version

  • Accepted version

Journal

Methods in Molecular Biology

ISSN

1064-3745

Publisher

Springer

Volume

2818

Page range

3-22

Book title

Meiosis. Methods in Molecular Biology

Place of publication

New York, USA

ISBN

9781071639054

Department affiliated with

  • Sussex Centre for Genome Damage Stability Publications

Institution

University of Sussex

Full text available

  • Yes

Peer reviewed?

  • Yes