染色质
染色质免疫沉淀
仿形(计算机编程)
计算生物学
表观遗传学
生物
化学
分子生物学
细胞生物学
DNA
基因
遗传学
生物化学
基因表达
DNA甲基化
发起人
作者
Xiaoyuan Tao,Shouli Feng,Ting Zhao,Xueying Guan
出处
期刊:Plant Methods
[Springer Nature]
日期:2020-08-31
卷期号:16 (1)
被引量:16
标识
DOI:10.1186/s13007-020-00664-8
摘要
Abstract Background In 2019, Kaya-Okur et al. reported on the cleavage under targets and tagmentation (CUT&Tag) technology for efficient profiling of epigenetically modified DNA fragments. It was used mainly for cultured cell lines and was especially effective for small samples and single cells. This strategy generated high-resolution and low-background-noise chromatin profiling data for epigenomic analysis. CUT&Tag is well suited to be used in plant cells, especially in tissues from which small samples are taken, such as ovules, anthers, and fibers. Results Here, we present a CUT&Tag protocol step by step using plant nuclei. In this protocol, we quantified the nuclei that can be used in each CUT&Tag reaction, and compared the efficiency of CUT&Tag with chromatin immunoprecipitation with sequencing (ChIP-seq) in the leaves of cotton. A general workflow for the bioinformatic analysis of CUT&Tag is also provided. Results indicated that, compared with ChIP-seq, the CUT&Tag procedure was faster and showed a higher-resolution, lower-background signal than did ChIP. Conclusion A CUT&Tag protocol has been refined for plant cells using intact nuclei that have been isolated.
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