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

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 approache...

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Detalles Bibliográficos
Autores principales: Li, Wangchun, Tim Wu, U, Cheng, Yu, Huang, Yanhao, Mao, Lipeng, Sun, Menghan, Qiu, Congling, Zhou, Lin, Gao, Lijuan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9388308/
https://www.ncbi.nlm.nih.gov/pubmed/36062065
http://dx.doi.org/10.1155/2022/8429207
Descripción
Sumario: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.