Epigenetic Application of ATAC-Seq Based on Tn5 Transposase Purification Technology.

IF 2.1 4区生物学 Q4 GENETICS & HEREDITY

Genetics research Pub Date : 2022-08-11 eCollection Date: 2022-01-01 DOI:10.1155/2022/8429207

Wangchun Li, U Tim Wu, Yu Cheng, Yanhao Huang, Lipeng Mao, Menghan Sun, Congling Qiu, Lin Zhou, Lijuan Gao

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引用次数: 0

Abstract

Background: Assays of transposase accessible chromatin sequencing (ATAC-seq) is an efficient assay to investigate chromatin accessibility, which depends on the activity of a robust Tn5 transposase to fragment the genome while cutting in the sequencing adapters.

Methods: We propose reliable approaches for purifying hyperactive Tn5 transposase by chitin magnetic bead sorting. Double-stranded DNA of J76 cells and 293T cells were digested and subjected to tagmentation as test samples with Tn5 transposase, and libraries were established and sequenced. Sequencing data was then analyzed for peak calling, GO enrichment, and motif analysis.

Results: We report a set of rapid, efficient, and low-cost methods for ATAC-seq library construction and data analysis, through large-scale and rapid sequencing. These methods can provide a reference for the study of epigenetic regulation of gene expression.

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基于Tn5转座酶纯化技术的ATAC-Seq的表观遗传应用

背景:转座酶可及性染色质测序(ATAC-seq)是一种研究染色质可及性的有效方法，它依赖于一个强大的Tn5转座酶的活性，在测序适配器中切割基因组片段。方法采用几丁质磁珠分选纯化高活性Tn5转座酶。将J76细胞和293T细胞的双链DNA作为测试样本，用Tn5转座酶进行酶切和标记，建立文库并测序。然后对测序数据进行峰召唤、氧化石墨烯富集和基序分析。结果:通过大规模快速测序，我们报告了一套快速、高效、低成本的ATAC-seq文库构建和数据分析方法。这些方法可为基因表达的表观遗传调控研究提供参考。

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来源期刊

Genetics research 生物-遗传学

自引率

6.70%

发文量

审稿时长

>12 weeks

期刊介绍： Genetics Research is a key forum for original research on all aspects of human and animal genetics, reporting key findings on genomes, genes, mutations and molecular interactions, extending out to developmental, evolutionary, and population genetics as well as ethical, legal and social aspects. Our aim is to lead to a better understanding of genetic processes in health and disease. The journal focuses on the use of new technologies, such as next generation sequencing together with bioinformatics analysis, to produce increasingly detailed views of how genes function in tissues and how these genes perform, individually or collectively, in normal development and disease aetiology. The journal publishes original work, review articles, short papers, computational studies, and novel methods and techniques in research covering humans and well-established genetic organisms. Key subject areas include medical genetics, genomics, human evolutionary and population genetics, bioinformatics, genetics of complex traits, molecular and developmental genetics, Evo-Devo, quantitative and statistical genetics, behavioural genetics and environmental genetics. The breadth and quality of research make the journal an invaluable resource for medical geneticists, molecular biologists, bioinformaticians and researchers involved in genetic basis of diseases, evolutionary and developmental studies.