Chromatin Immunoprecipitation Experiments from Whole Drosophila Embryos or Larval Imaginal Discs

Vincent Loubiere; Anna Delest; Bernd Schuettengruber; Anne-Marie Martinez; Giacomo Cavalli

doi:10.21769/BioProtoc.2327

Chromatin Immunoprecipitation Experiments from Whole Drosophila Embryos or Larval Imaginal Discs

Bio Protoc. 2017 Jun 5;7(11):e2327. doi: 10.21769/BioProtoc.2327.

Authors

Vincent Loubiere^{1

2}, Anna Delest³, Bernd Schuettengruber¹, Anne-Marie Martinez^{1

2}, Giacomo Cavalli¹

Affiliations

¹ Institute of Human Genetics, UMR9002 CNRS-UM, 141 Rue de la Cardonille, 34396 Montpellier Cedex 5, France.
² University of Montpellier, 163 Rue Auguste Broussonnet, 34090 Montpellier, France.
³ Centre de Recherche en Cancérologie de Lyon, INSERM U1052, 151 cours Albert Thomas, 69003 Lyon, France.

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.

Keywords: ChIP; Drosophila; Embryo; Epigenetic mark; Imaginal disc; Transcription factor.