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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...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Hindawi
2022
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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 |
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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 |
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