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SALP, a new single-stranded DNA library preparation method especially useful for the high-throughput characterization of chromatin openness states

BACKGROUND: Next-generation sequencing (NGS) is fundamental to the current biological and biomedical research. Construction of sequencing library is a key step of NGS. Therefore, various library construction methods have been explored. However, the current methods are still limited by some shortcomi...

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Detalles Bibliográficos
Autores principales: Wu, Jian, Dai, Wei, Wu, Lin, Wang, Jinke
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5811972/
https://www.ncbi.nlm.nih.gov/pubmed/29439663
http://dx.doi.org/10.1186/s12864-018-4530-3
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author Wu, Jian
Dai, Wei
Wu, Lin
Wang, Jinke
author_facet Wu, Jian
Dai, Wei
Wu, Lin
Wang, Jinke
author_sort Wu, Jian
collection PubMed
description BACKGROUND: Next-generation sequencing (NGS) is fundamental to the current biological and biomedical research. Construction of sequencing library is a key step of NGS. Therefore, various library construction methods have been explored. However, the current methods are still limited by some shortcomings. RESULTS: This study developed a new NGS library construction method, Single strand Adaptor Library Preparation (SALP), by using a novel single strand adaptor (SSA). SSA is a double-stranded oligonucleotide with a 3′ overhang of 3 random nucleotides, which can be efficiently ligated to the 3′ end of single strand DNA by T4 DNA ligase. SALP can be started with any denatured DNA fragments such as those sheared by Tn5 tagmentation, enzyme digestion and sonication. When started with Tn5-tagmented chromatin, SALP can overcome a key limitation of ATAC-seq and become a high-throughput NGS library construction method, SALP-seq, which can be used to comparatively characterize the chromatin openness state of multiple cells unbiasly. In this way, this study successfully characterized the comparative chromatin openness states of four different cell lines, including GM12878, HepG2, HeLa and 293T, with SALP-seq. Similarly, this study also successfully characterized the chromatin openness states of HepG2 cells with SALP-seq by using 10(5) to 500 cells. CONCLUSIONS: This study developed a new NGS library construction method, SALP, by using a novel kind of single strand adaptor (SSA), which should has wide applications in the future due to its unique performance. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12864-018-4530-3) contains supplementary material, which is available to authorized users.
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spelling pubmed-58119722018-02-15 SALP, a new single-stranded DNA library preparation method especially useful for the high-throughput characterization of chromatin openness states Wu, Jian Dai, Wei Wu, Lin Wang, Jinke BMC Genomics Methodology Article BACKGROUND: Next-generation sequencing (NGS) is fundamental to the current biological and biomedical research. Construction of sequencing library is a key step of NGS. Therefore, various library construction methods have been explored. However, the current methods are still limited by some shortcomings. RESULTS: This study developed a new NGS library construction method, Single strand Adaptor Library Preparation (SALP), by using a novel single strand adaptor (SSA). SSA is a double-stranded oligonucleotide with a 3′ overhang of 3 random nucleotides, which can be efficiently ligated to the 3′ end of single strand DNA by T4 DNA ligase. SALP can be started with any denatured DNA fragments such as those sheared by Tn5 tagmentation, enzyme digestion and sonication. When started with Tn5-tagmented chromatin, SALP can overcome a key limitation of ATAC-seq and become a high-throughput NGS library construction method, SALP-seq, which can be used to comparatively characterize the chromatin openness state of multiple cells unbiasly. In this way, this study successfully characterized the comparative chromatin openness states of four different cell lines, including GM12878, HepG2, HeLa and 293T, with SALP-seq. Similarly, this study also successfully characterized the chromatin openness states of HepG2 cells with SALP-seq by using 10(5) to 500 cells. CONCLUSIONS: This study developed a new NGS library construction method, SALP, by using a novel kind of single strand adaptor (SSA), which should has wide applications in the future due to its unique performance. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12864-018-4530-3) contains supplementary material, which is available to authorized users. BioMed Central 2018-02-13 /pmc/articles/PMC5811972/ /pubmed/29439663 http://dx.doi.org/10.1186/s12864-018-4530-3 Text en © The Author(s). 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Methodology Article
Wu, Jian
Dai, Wei
Wu, Lin
Wang, Jinke
SALP, a new single-stranded DNA library preparation method especially useful for the high-throughput characterization of chromatin openness states
title SALP, a new single-stranded DNA library preparation method especially useful for the high-throughput characterization of chromatin openness states
title_full SALP, a new single-stranded DNA library preparation method especially useful for the high-throughput characterization of chromatin openness states
title_fullStr SALP, a new single-stranded DNA library preparation method especially useful for the high-throughput characterization of chromatin openness states
title_full_unstemmed SALP, a new single-stranded DNA library preparation method especially useful for the high-throughput characterization of chromatin openness states
title_short SALP, a new single-stranded DNA library preparation method especially useful for the high-throughput characterization of chromatin openness states
title_sort salp, a new single-stranded dna library preparation method especially useful for the high-throughput characterization of chromatin openness states
topic Methodology Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5811972/
https://www.ncbi.nlm.nih.gov/pubmed/29439663
http://dx.doi.org/10.1186/s12864-018-4530-3
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