title: Chromatin Immunoprecipitation for Analyzing Transcription Factor Binding and Histone Modifications in Drosophila.
abstract: Chromatin immunoprecipitation followed by next-generation sequencing (ChIP-seq) is an invaluable technique to assess transcription factor binding and histone modifications in a genome-wide manner, an essential step towards understanding the mechanisms that govern embryonic development. Here, we provide a detailed protocol for all steps involved in generating a ChIP-seq library, starting from embryo collection, fixation, chromatin preparation, immunoprecipitation, and finally library preparation. The protocol is optimized for Drosophila embryos, but can be easily adapted for any model organism. The resulting library is suitable for sequencing on an Illumina HiSeq or MiSeq platform.
authors.affiliation: European Molecular Biology Laboratory, Genome Biology Unit, Heidelberg, D-69117, Germany.
authors.forename: Yad
authors.initials: Y
authors.lastname: Ghavi-Helm
authors.affiliation: European Molecular Biology Laboratory, Genome Biology Unit, Heidelberg, D-69117, Germany.
authors.forename: Bingqing
authors.initials: B
authors.lastname: Zhao
authors.affiliation: European Molecular Biology Laboratory, Genome Biology Unit, Heidelberg, D-69117, Germany. furlong@embl.de.
authors.forename: Eileen E M
authors.initials: EE
authors.lastname: Furlong
mesh_terms: D000818:Animals; D002843:Chromatin / Q000737:chemistry / Q000378:metabolism; D042002:Chromatin Assembly and Disassembly; D047369:Chromatin Immunoprecipitation / Q000379:methods; D004331:Drosophila melanogaster / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D004625:Embryo, Nonmammalian; D015723:Gene Library; D016678:Genome; D059014:High-Throughput Nucleotide Sequencing; D006657:Histones / Q000235:genetics* / Q000378:metabolism; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D015398:Signal Transduction; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D002843:Chromatin; D006657:Histones; D014157:Transcription Factors
keywords: ChIP-seq; Chromatin immunoprecipitation; Development; Drosophila; Next-generation sequencing; Transcription