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

Author:

Li Wangchun1,Tim Wu U2,Cheng Yu34,Huang Yanhao34,Mao Lipeng34,Sun Menghan34,Qiu Congling4,Zhou Lin5ORCID,Gao Lijuan34ORCID

Affiliation:

1. Department of Critical Care Medicine, Shunde Hospital Affiliated of Jinan University, 528305 Foshan, China

2. Meng Yi Center Limited, Macau 999078, China

3. Institute of Geriatric Immunology, School of Medicine, Jinan University, 510632 Guangzhou, China

4. Department of Microbiology and Immunology, School of Medicine, Jinan University, Guangzhou, China

5. Nanjing Maternity and Child Health Care Hospital, Women’s Hospital of Nanjing Medical University, No. 123, Tianfeixiang, Mochou Rd., Nanjing 210004, China

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.

Funder

Medical Science and Technology Foundation of Guangdong Province

Publisher

Hindawi Limited

Subject

Genetics,General Medicine

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