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Meiosis

Volume 557 of the series Methods in Molecular Biology pp 183-195

Date:

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

  • Matthew J. NealeAffiliated withHoward Hughes Medical Institute and Molecular Biology Program, Memorial Sloan-Kettering Cancer Center
  • , Scott KeeneyAffiliated withMRC Genome Damage and Stability Centre, University of Sussex, Falmer

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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