Titlebook: DNA Recombination; Methods and Protocol Hideo Tsubouchi Book 2011 Springer Science+Business Media, LLC 2011 DNA double-strand break.DNA mic

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Methods to Study Mitotic Homologous Recombination and Genome Stabilityeave recombinogenic substrates such as single-stranded DNA gaps. Other types of damages result in general genome instability such as chromosome loss, chromosome fragmentation, and chromosome rearrangements. The genome is kept intact through recombination, repair, replication, checkpoints, and chromo

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Characterizing Resection at Random and Unique Chromosome Double-Strand Breaks and Telomere Endsrminant in choice of repair pathways and eventual genome stability. Current techniques for examining resection are restricted to model in vivo systems with defined substrates (i.e., HO-endonuclease targets). We present here a robust assay that can analyze not only the resection of site-specific DSBs

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Genome-Wide Detection of Meiotic DNA Double-Strand Break Hotspots Using Single-Stranded DNAome segregation in most eukaryotes. This chapter describes a straightforward microarray-based approach to measure the genome-wide distribution of meiotic DSBs by detecting the single-stranded DNA (ssDNA) that transiently accumulates at DSB sites during recombination. The protocol outlined here has b

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Detection of Covalent DNA-Bound Spo11 and Topoisomerase Complexes DNA break, the topoisomerase remains covalently linked to the DNA and removes itself when the break is re-ligated. While the lifetime of a covalent topoisomerase–DNA complex is usually short, several clinically important cancer drugs kill cancer cells by inhibiting the removal of covalently linked

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Analysis of Meiotic Recombination Intermediates by Two-Dimensional Gel Electrophoresisrmed via several branched intermediates, including single end invasions and double Holliday junctions. Two-dimensional gel electrophoresis provides a sensitive and specific approach for detecting branched recombination intermediates, determining their genetic requirements, and enriching intermediate

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Mapping of Crossover Sites Using DNA Microarraysysical bridges between homologs that provide the appropriate tension to properly align chromosomes on the meiosis I spindle. Homolog pairs that fail to cross over can result in meiosis I nondisjunction, leading to aneuploid gametes. Therefore, the number and distribution of crossovers are tightly re

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