Chromatin Immunoprecipitation Experiments from Whole Drosophila Embryos or Larval Imaginal Discs

整个果蝇胚胎或幼虫成虫盘的染色质免疫沉淀实验

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作者:Vincent Loubiere, Anna Delest, Bernd Schuettengruber, Anne-Marie Martinez, Giacomo Cavalli

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.

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