| Literature DB >> 34541088 |
Vincent Loubiere1,2, Anna Delest3, Bernd Schuettengruber1, Anne-Marie Martinez1,2, Giacomo Cavalli1.
Abstract
Chromatin Immunoprecipitation coupled either to qPCR (qChIP) or high-throughput sequencing (ChIP-Seq) has been extensively used in the last decades to identify the DNA binding sites of transcription factors or the localization of various histone marks along the genome. The ChIP experiment generally includes 7 steps: collection of biological samples (A), cross-linking proteins to DNA (B), chromatin isolation and fragmentation by sonication (C), sonication test (D), immunoprecipitation with antibodies against the protein or the histone mark of interest (E), DNA recovery (E), identification of factor-associated DNA sequences by PCR or sequencing (F). The protocol described here can readily be used for ChIP-seq and ChIP-qPCR experiments. The entire procedure, describing experimental setup conditions to optimize assays in intact Drosophila tissues, can be completed within four days.Entities:
Keywords: ChIP; Drosophila; Embryo; Epigenetic mark; Imaginal disc; Transcription factor
Year: 2017 PMID: 34541088 PMCID: PMC8410254 DOI: 10.21769/BioProtoc.2327
Source DB: PubMed Journal: Bio Protoc ISSN: 2331-8325