Gentsch George E, Smith James C
The Francis Crick Institute, Developmental Biology Laboratory, 1 Midland Road, London, NW1 1AT, UK.
Methods Mol Biol. 2017;1507:23-42. doi: 10.1007/978-1-4939-6518-2_3.
Chromatin immunoprecipitation followed by next-generation sequencing (ChIP-seq) has become a powerful tool to acquire a precise and genome-wide snapshot of many chromatin features in vivo. These chromatin profiles are obtained by immunoprecipitation of cross-linked chromatin fragments to enrich the feature of interest. Sequencing and aligning the underlying DNA sequences to the genome make it possible to virtually reconstruct the global distribution of most chromatin features. We present here recent improvements to the ChIP-seq protocol by means of Xenopus embryos to prepare high-complexity DNA libraries from small amounts of biological material. This approach allows researchers to explore the landscape of chromatin regulators and states in early vertebrate embryos or in any biological entity with small numbers of cells.
染色质免疫沉淀结合新一代测序技术(ChIP-seq)已成为一种强大的工具,可在体内获取许多染色质特征的精确全基因组快照。这些染色质图谱是通过对交联的染色质片段进行免疫沉淀以富集感兴趣的特征而获得的。对潜在的DNA序列进行测序并与基因组比对,使得几乎可以重建大多数染色质特征的全局分布。我们在此展示了通过非洲爪蟾胚胎对ChIP-seq方案进行的最新改进,以便从少量生物材料中制备高复杂性DNA文库。这种方法使研究人员能够探索早期脊椎动物胚胎或任何细胞数量较少的生物实体中的染色质调节因子和状态。