van Mourik Hilda, Muiño Jose M, Pajoro Alice, Angenent Gerco C, Kaufmann Kerstin
Laboratory of Molecular Biology, Wageningen University, 6708PB, Wageningen, The Netherlands.
Methods Mol Biol. 2015;1284:93-121. doi: 10.1007/978-1-4939-2444-8_5.
Chromatin immunoprecipitation followed by next-generation sequencing (ChIP-seq) is a powerful technique for genome-wide identification of in vivo binding sites of DNA-binding proteins. The technique had been used to study many DNA-binding proteins in a broad variety of species. The basis of the ChIP-seq technique is the ability to covalently cross-link DNA and proteins that are located in very close proximity. This allows the use of an antibody against the (tagged) protein of interest to specifically enrich DNA-fragments bound by this protein. ChIP-seq can be performed using antibodies against the native protein or against tagged proteins. Using a specific antibody against a tag to immunoprecipitate tagged proteins eliminates the need for a specific antibody against the native protein and allows more experimental flexibility. In this chapter we present a complete workflow for experimental procedure and bioinformatic analysis that allows wet-lab biologists to perform and analyze ChIP-seq experiments.
染色质免疫沉淀结合下一代测序（ChIP-seq）是一种用于全基因组范围内鉴定DNA结合蛋白体内结合位点的强大技术。该技术已被用于研究多种物种中的许多DNA结合蛋白。ChIP-seq技术的基础是能够共价交联非常接近的DNA和蛋白质。这使得可以使用针对感兴趣的（标记）蛋白质的抗体来特异性富集该蛋白质结合的DNA片段。ChIP-seq可以使用针对天然蛋白质或标记蛋白质的抗体来进行。使用针对标签的特异性抗体免疫沉淀标记蛋白质消除了对针对天然蛋白质的特异性抗体的需求，并提供了更大的实验灵活性。在本章中，我们介绍了一个完整的实验流程和生物信息学分析工作流程，使湿实验室生物学家能够进行和分析ChIP-seq实验。
