Meiosis pp 183-195 | Cite as

End-Labeling and Analysis of Spo11-Oligonucleotide Complexes in Saccharomyces cerevisiae

  • Matthew J. Neale
  • Scott Keeney
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 557)

Abstract

During meiosis Spo11 catalyzes the formation of DNA double-strand breaks, becoming covalently attached to the 5′ ends on both sides of the break during this process. Spo11 is removed from the DSB by single-stranded endonucleolytic cleavage flanking the DSB, liberating a short-lived species consisting of Spo11 protein covalently linked to a short oligonucleotide. The method presented here details how to detect these Spo11-oligo complexes in extracts made from meiotic yeast cells.

Key words

Spo11 meiosis DNA double-strand break end-labeling terminal transferase 

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Matthew J. Neale
    • 2
  • Scott Keeney
    • 1
  1. 1.MRC Genome Damage and Stability CentreUniversity of Sussex, FalmerUK
  2. 2.Howard Hughes Medical Institute and Molecular Biology Program, Memorial Sloan-Kettering Cancer CenterNew YorkUSA

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