Abstract
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.
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References
Bellve, A.R. (1993) Purification, culture, and fractionation of spermatogenic cells. Methods Enzymol. 225, 84–113.
Neale, M.J., Pan, J., Keeney, S. (2005) Endonucleolytic processing of covalent protein-linked DNA double-strand breaks. Nature 436, 1053–1057.
Baudat, F., de Massy, B. (2005) Regulating double-stranded DNA break repair towards crossover or non-crossover during mammalian meiosis. Chromosome Res. 15, 565–77.
Keeney, S., Baudat, F., Angeles, M., Zhou, Z.H., Copeland, N.G., Jenkins, N.A., Manova, K., Jasin, M. (1999) A mouse homolog of the Saccharomyces cerevisiae meiotic recombination DNA transesterase Spo11p. Genomics 61, 170–182.
Romanienko, P.J., Camerini-Otero, R.D. (1999) Cloning, characterization, and localization of mouse and human SPO11. Genomics 61, 156–169.
Sambrook, J., Russel, D.W. (2001) Molecular Cloning. Chapter 12. Protocols 8, 11, and 12. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Pan, J., Keeney, S. (2009). Detection of SPO11-Oligonucleotide Complexes from Mouse Testes. In: Keeney, S. (eds) Meiosis. Methods in Molecular Biology, vol 557. Humana Press. https://doi.org/10.1007/978-1-59745-527-5_13
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DOI: https://doi.org/10.1007/978-1-59745-527-5_13
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Publisher Name: Humana Press
Print ISBN: 978-1-934115-66-4
Online ISBN: 978-1-59745-527-5
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