Chromatin Immunoprecipitation Experiments from Whole Drosophila Embryos or Larval Imaginal Discs

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...

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Bibliographic Details
Published inBio-protocol Vol. 7; no. 11; p. e2327
Main Authors Loubiere, Vincent, Delest, Anna, Schuettengruber, Bernd, Martinez, Anne-Marie, Cavalli, Giacomo
Format Journal Article
LanguageEnglish
Published United States Bio-Protocol 05.06.2017
Bio-protocol LLC
Subjects
Biology
Methods
Embryo
Epigenetic mark
Imaginal disc
ChIP
Transcription factor
Drosophila
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Summary: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 tissues, can be completed within four days.
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ISSN:2331-8325
2331-8325
DOI:10.21769/BioProtoc.2327