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使用转座酶辅助片段化方法对K562细胞和神经母细胞的有限起始材料进行染色质免疫沉淀测序(ChIP-Seq)

ChIP-Seq from Limited Starting Material of K562 Cells and Neuroblasts Using Tagmentation Assisted Fragmentation Approach.

作者信息

Akhtar Junaid, More Piyush, Albrecht Steffen

机构信息

Institute of Developmental Biology and Neurobiology, University of Mainz, Hanns-Dieter-Hsch Weg 15, 55128 Mainz, Germany.

Department of Pharmacology, University Medical Center, Johannes Gutenberg University of Mainz, 55131 Mainz, Germany.

出版信息

Bio Protoc. 2020 Feb 20;10(4):e3520. doi: 10.21769/BioProtoc.3520.

Abstract

Chromatin immunoprecipitation is extensively used to investigate the epigenetic profile and transcription factor binding sites in the genome. However, when the starting material is limited, the conventional ChIP-Seq approach cannot be implemented. This protocol describes a method that can be used to generate the chromatin profiles from as low as 100 human or 1,000 cells. The method employs tagmentation to fragment the chromatin with concomitant addition of sequencing adaptors. The method generates datasets with high signal to noise ratio and can be subjected to standard tools for ChIP-Seq analysis.

摘要

染色质免疫沉淀技术被广泛用于研究基因组中的表观遗传图谱和转录因子结合位点。然而,当起始材料有限时,传统的ChIP-Seq方法就无法实施。本方案描述了一种方法,该方法可用于从低至100个人类细胞或1000个细胞中生成染色质图谱。该方法采用转座酶标签法对染色质进行片段化,并同时添加测序接头。该方法生成的数据集具有高信噪比,并且可以使用ChIP-Seq分析的标准工具进行分析。

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