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CC-seq: Nucleotide-resolution mapping of Spo11 DNA double-strand breaks in S. cerevisiae cells

George G. B. BrowniD, William H. GittensiD, Rachal M. AllisoniD, Antony W. OliveriD, Matthew J. NealeiD

Abstract

i.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 synchronised Saccharomyces cerevisiae meiotic cells.

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