An Integrated and Semiautomated Microscaled Approach to Profile Cis-Regulatory Elements by Histone Modification ChIP-Seq for Large-Scale Epigenetic Studies
An Integrated and Semiautomated Microscaled Approach to Profile Cis-Regulatory Elements by Histone Modification ChIP-Seq for Large-Scale Epigenetic Studies
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DOI：
10.1007/978-1-4939-7896-0_22
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发表时间：
2018-01-01
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影响因子：
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通讯作者：
Seumois, Gregory
中科院分区：
文献类型：
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作者：
Jankeel, Diana Youhanna;Cayford, Justin;Seumois, Gregory
染色质免疫沉淀测序（ChIP-Seq）是绘制组蛋白修饰图和鉴定整个基因组顺式调控DNA元件的首选方法。已经描述了多种方法来提高文库制备的效率并减少动手时间以及成本。本文介绍了一批48-96个小细胞样本的细胞固定、染色质剪切、免疫沉淀和测序文库制备的详细步骤。该方案实现了一个半自动化平台，以减少技术变异性，提高信噪比，并减少动手时间，从而允许对细胞数量有限的临床样本进行大规模表观遗传学研究。
Chromatin immunoprecipitation followed by sequencing (ChIP-Seq) is the preferred approach to map histone modifications and identify cis-regulatory DNA elements throughout the genome. Multiple methods have been described to increase the efficiency of library preparation and to reduce hands-on time as well as costs. This review describes detailed steps to perform cell fixation, chromatin shearing, immunoprecipitation, and sequencing library preparation for a batch of 48-96 samples with small cell numbers. The protocol implements a semiautomated platform to reduce technical variability and improve signal-to-noise ratio as well as reduce hands-on time, thus allowing large-scale epigenetic studies of clinical samples with limited cell numbers.