Volume 557 of the series Methods in Molecular Biology pp 197-207


Detection of SPO11-Oligonucleotide Complexes from Mouse Testes

  • Jing PanAffiliated withMolecular Biology Program, Memorial Sloan-Kettering Cancer Center
  • , Scott KeeneyAffiliated withHoward Hughes Medical Institute and Molecular Biology Program, Memorial Sloan-Kettering Cancer Center

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The SPO11 protein generates programmed DNA double-strand breaks (DSBs) that initiate meiotic recombination. Endonucleolytic cleavage 3′ to the DSB sites releases SPO11 from DNA, leaving SPO11 covalently associated with an oligonucleotide. This chapter describes detection of the release product, SPO11-oligonucleotide complexes, from mouse testis lysates. The method for determining the size of SPO11-associated oligonucleotides is also provided.

Key words

SPO11 mouse testes terminal transferase DNA double-strand break