Cargando…

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...

Descripción completa

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
_version_ 1784770198476161024
author Li, Wangchun
Tim Wu, U
Cheng, Yu
Huang, Yanhao
Mao, Lipeng
Sun, Menghan
Qiu, Congling
Zhou, Lin
Gao, Lijuan
author_facet Li, Wangchun
Tim Wu, U
Cheng, Yu
Huang, Yanhao
Mao, Lipeng
Sun, Menghan
Qiu, Congling
Zhou, Lin
Gao, Lijuan
author_sort Li, Wangchun
collection PubMed
description 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.
format Online
Article
Text
id pubmed-9388308
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Hindawi
record_format MEDLINE/PubMed
spelling pubmed-93883082022-09-02 Epigenetic Application of ATAC-Seq Based on Tn5 Transposase Purification Technology Li, Wangchun Tim Wu, U Cheng, Yu Huang, Yanhao Mao, Lipeng Sun, Menghan Qiu, Congling Zhou, Lin Gao, Lijuan Genet Res (Camb) Research Article 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. Hindawi 2022-08-11 /pmc/articles/PMC9388308/ /pubmed/36062065 http://dx.doi.org/10.1155/2022/8429207 Text en Copyright © 2022 Wangchun Li et al. https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Li, Wangchun
Tim Wu, U
Cheng, Yu
Huang, Yanhao
Mao, Lipeng
Sun, Menghan
Qiu, Congling
Zhou, Lin
Gao, Lijuan
Epigenetic Application of ATAC-Seq Based on Tn5 Transposase Purification Technology
title Epigenetic Application of ATAC-Seq Based on Tn5 Transposase Purification Technology
title_full Epigenetic Application of ATAC-Seq Based on Tn5 Transposase Purification Technology
title_fullStr Epigenetic Application of ATAC-Seq Based on Tn5 Transposase Purification Technology
title_full_unstemmed Epigenetic Application of ATAC-Seq Based on Tn5 Transposase Purification Technology
title_short Epigenetic Application of ATAC-Seq Based on Tn5 Transposase Purification Technology
title_sort epigenetic application of atac-seq based on tn5 transposase purification technology
topic Research Article
url 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
work_keys_str_mv AT liwangchun epigeneticapplicationofatacseqbasedontn5transposasepurificationtechnology
AT timwuu epigeneticapplicationofatacseqbasedontn5transposasepurificationtechnology
AT chengyu epigeneticapplicationofatacseqbasedontn5transposasepurificationtechnology
AT huangyanhao epigeneticapplicationofatacseqbasedontn5transposasepurificationtechnology
AT maolipeng epigeneticapplicationofatacseqbasedontn5transposasepurificationtechnology
AT sunmenghan epigeneticapplicationofatacseqbasedontn5transposasepurificationtechnology
AT qiucongling epigeneticapplicationofatacseqbasedontn5transposasepurificationtechnology
AT zhoulin epigeneticapplicationofatacseqbasedontn5transposasepurificationtechnology
AT gaolijuan epigeneticapplicationofatacseqbasedontn5transposasepurificationtechnology