Berguet Geoffrey, Hendrickx Jan, Sabatel Celine, Laczik Miklos, Squazzo Sharon, Mazon Pelaez Ignacio, Saxena Rini, Pendeville Helene, Poncelet Dominique
Diagenode S.A.
Diagenode Inc.
J Vis Exp. 2014 Dec 10(94):52150. doi: 10.3791/52150.
Chromatin immunoprecipitation followed by next generation sequencing (ChIP-seq) is a technique of choice for studying protein-DNA interactions. ChIP-seq has been used for mapping protein-DNA interactions and allocating histones modifications. The procedure is tedious and time consuming, and one of the major limitations is the requirement for high amounts of starting material, usually millions of cells. Automation of chromatin immunoprecipitation assays is possible when the procedure is based on the use of magnetic beads. Successful automated protocols of chromatin immunoprecipitation and library preparation have been specifically designed on a commercially available robotic liquid handling system dedicated mainly to automate epigenetic assays. First, validation of automated ChIP-seq assays using antibodies directed against various histone modifications was shown, followed by optimization of the automated protocols to perform chromatin immunoprecipitation and library preparation starting with low cell numbers. The goal of these experiments is to provide a valuable tool for future epigenetic analysis of specific cell types, sub-populations, and biopsy samples.
染色质免疫沉淀结合下一代测序（ChIP-seq）是研究蛋白质-DNA相互作用的首选技术。ChIP-seq已被用于绘制蛋白质-DNA相互作用图谱和分配组蛋白修饰。该过程繁琐且耗时，主要限制之一是需要大量起始材料，通常为数百万个细胞。当染色质免疫沉淀测定程序基于磁珠的使用时，实现自动化是可能的。专门针对主要用于自动化表观遗传学测定的市售机器人液体处理系统，成功设计了染色质免疫沉淀和文库制备的自动化方案。首先，展示了使用针对各种组蛋白修饰的抗体对自动化ChIP-seq测定的验证，随后优化了自动化方案，以便从低细胞数量开始进行染色质免疫沉淀和文库制备。这些实验的目的是为未来对特定细胞类型、亚群和活检样本的表观遗传学分析提供有价值的工具。
