End-Labeling and Analysis of Spo11-Oligonucleotide Complexes in Saccharomyces cerevisiae
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 wordsSpo11 meiosis DNA double-strand break end-labeling terminal transferase