Cell type-specific chromatin immunoprecipitation from multicellular complex samples using BiTS-ChIP

摘要: This protocol describes the batch isolation of tissue-specific chromatin for immunoprecipitation (BiTS-ChIP) analysis histone modifications, transcription factor binding, or polymerase occupancy within context a multicellular organism tissue. Embryos expressing cell type-specific nuclear marker are formaldehyde cross-linked and then subjected to dissociation. Fixed nuclei isolated sorted using FACS on basis marker. Tissue-specific is extracted, sheared by sonication used ChIP-seq other analyses. The key advantages this method covalent cross-linking before embryo dissociation, which preserves transcriptional context, use nuclei, yielding very high purity. has been optimized Drosophila, but with minor modifications should be applicable any model system. full protocol, including sorting, generation sequencing libraries, can completed 5 d.