Ghavi-Helm Yad, Zhao Bingqing, Furlong Eileen E M
European Molecular Biology Laboratory, Genome Biology Unit, Heidelberg, D-69117, Germany.
Methods Mol Biol. 2016;1478:263-277. doi: 10.1007/978-1-4939-6371-3_16.
Chromatin immunoprecipitation followed by next-generation sequencing (ChIP-seq) is an invaluable technique to assess transcription factor binding and histone modifications in a genome-wide manner, an essential step towards understanding the mechanisms that govern embryonic development. Here, we provide a detailed protocol for all steps involved in generating a ChIP-seq library, starting from embryo collection, fixation, chromatin preparation, immunoprecipitation, and finally library preparation. The protocol is optimized for Drosophila embryos, but can be easily adapted for any model organism. The resulting library is suitable for sequencing on an Illumina HiSeq or MiSeq platform.
染色质免疫沉淀结合新一代测序技术(ChIP-seq)是一种极为重要的技术,可在全基因组范围内评估转录因子结合和组蛋白修饰,这是理解胚胎发育调控机制的关键步骤。在此,我们提供了一个详细的方案,涵盖生成ChIP-seq文库所涉及的所有步骤,从胚胎收集、固定、染色质制备、免疫沉淀,直至最终的文库制备。该方案针对果蝇胚胎进行了优化,但可轻松适用于任何模式生物。所得文库适用于在Illumina HiSeq或MiSeq平台上进行测序。