Methods in Molecular Biology Volume 557, 2009, pp 183-195
Date: 25 May 2009

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

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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.