title: Mapping Recombination Initiation Sites Using Chromatin Immunoprecipitation.
abstract: Genome-wide maps of recombination sites provide valuable information not only on the recombination pathway itself but also facilitate the understanding of genome dynamics and evolution. Here, we describe a chromatin immunoprecipitation (ChIP) protocol to map the sites of recombination initiation in plants with maize used as an example. ChIP is a method that allows identification of chromosomal sites occupied by specific proteins. Our protocol utilizes RAD51, a protein involved in repair of double-strand breaks (DSBs) that initiate meiotic recombination, to identify DSB formation hotspots. Chromatin is extracted from meiotic flowers, sheared and enriched in fragments bound to RAD51. Genomic location of the protein is then identified by next-generation sequencing. This protocol can also be used in other species of plants, animals, and fungi.
authors: National Maize Improvement Center of China, Beijing Key Laboratory of Crop Genetic Improvement, China Agricultural University, Beijing, 100083, China.
authors: Yan
authors: Y
authors: He
authors: Institute of Biotechnology, Biotechnology Resource Center and Section of Plant Biology in School of IntegrativePlant Science, Cornell University, Ithaca, NY, 14853, USA.
authors: Minghui
authors: M
authors: Wang
authors: Institute of Biotechnology, Biotechnology Resource Center, Cornell University, Ithaca, NY, 14853, USA.
authors: Qi
authors: Q
authors: Sun
authors: Section of Plant Biology, School of Integrative Plant Science, Cornell University, Ithaca, NY, 14853, USA. wp45@cornell.edu.
authors: Wojciech P
authors: WP
authors: Pawlowski
mesh_terms: D047369:Chromatin Immunoprecipitation / Q000379:methods*; D002874:Chromosome Mapping / Q000379:methods*; D032461:Chromosomes, Plant; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D051135:Rad51 Recombinase / Q000235:genetics; D011995:Recombination, Genetic; D024363:Transcription Initiation Site; D003313:Zea mays / Q000235:genetics*
chemical_list: D051135:Rad51 Recombinase
keywords: Antibody; Chromatin; Chromosomes; Double-strand breaks (DSBs); Immunoprecipitation; Maize; Recombination