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<title lang=en>Sensitive mapping of recombination hotspots using sequencing-based detection of ssDNA.</title>
<creator>Khil, Pavel P</creator>
<creator>Smagulova, Fatima</creator>
<creator>Brick, Kevin M</creator>
<creator>Camerini-Otero, R Daniel</creator>
<creator>Petukhova, Galina V</creator>
<contributor>Genetics and Biochemistry Branch ; National Institute of Health (NIH) - National Institute of Diabetes, Digestive and Kidney Diseases</contributor>
<contributor>Department of Biochemistry and Molecular Biology ; Uniformed Services University of the Health Sciences</contributor>
<contributor>Institut de recherche, santé, environnement et travail [Rennes] (Irset) ; Université d'Angers (UA) - Université des Antilles et de la Guyane (UAG) - Université de Rennes 1 (UR1) - École des Hautes Études en Santé Publique [EHESP] (EHESP) - Institut National de la Santé et de la Recherche Médicale (INSERM) - Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )</contributor>
<contributor>NIDDK Intramural Research Program; Basil O'Connor Starter Scholar Research Award Grant No. 5-FY07-667 from the March of Dimes Foundation (G.V.P.); NIH grant 1R01GM084104-01A1 from NIGMS (G.V.P.); New Investigator Start-up Grants FS71HU, R071HU ; CS71HU from USUHS (G.V.P.)</contributor>
<description>International audience</description>
<source>ISSN: 1088-9051</source>
<source>EISSN: 1549-5469</source>
<source>Genome Research</source>
<publisher>Cold Spring Harbor Laboratory Press</publisher>
<identifier>hal-00877675</identifier>
<identifier>https://hal.archives-ouvertes.fr/hal-00877675</identifier>
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<source>Genome Research, Cold Spring Harbor Laboratory Press, 2012, 22 (5), pp.957-65. 〈10.1101/gr.130583.111〉</source>
<identifier>DOI : 10.1101/gr.130583.111</identifier>
<relation>info:eu-repo/semantics/altIdentifier/doi/10.1101/gr.130583.111</relation>
<identifier>PUBMED : 22367190</identifier>
<relation>info:eu-repo/semantics/altIdentifier/pmid/22367190</relation>
<language>en</language>
<subject>[SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology</subject>
<type>info:eu-repo/semantics/article</type>
<type>Journal articles</type>
<description lang=en>Meiotic DNA double-stranded breaks (DSBs) initiate genetic recombination in discrete areas of the genome called recombination hotspots. DSBs can be directly mapped using chromatin immunoprecipitation followed by sequencing (ChIP-seq). Nevertheless, the genome-wide mapping of recombination hotspots in mammals is still a challenge due to the low frequency of recombination, high heterogeneity of the germ cell population, and the relatively low efficiency of ChIP. To overcome these limitations we have developed a novel method--single-stranded DNA (ssDNA) sequencing (SSDS)--that specifically detects protein-bound single-stranded DNA at DSB ends. SSDS comprises a computational framework for the specific detection of ssDNA-derived reads in a sequencing library and a new library preparation procedure for the enrichment of fragments originating from ssDNA. The use of our technique reduces the nonspecific double-stranded DNA (dsDNA) background >10-fold. Our method can be extended to other systems where the identification of ssDNA or DSBs is desired.</description>
<date>2012-05</date>
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