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INC-Seq: accurate single molecule reads using nanopore sequencing

BACKGROUND: Nanopore sequencing provides a rapid, cheap and portable real-time sequencing platform with the potential to revolutionize genomics. However, several applications are limited by relatively high single-read error rates (>10 %), including RNA-seq, haplotype sequencing and 16S sequencing...

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Autores principales: Li, Chenhao, Chng, Kern Rei, Boey, Esther Jia Hui, Ng, Amanda Hui Qi, Wilm, Andreas, Nagarajan, Niranjan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4970289/
https://www.ncbi.nlm.nih.gov/pubmed/27485345
http://dx.doi.org/10.1186/s13742-016-0140-7
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author Li, Chenhao
Chng, Kern Rei
Boey, Esther Jia Hui
Ng, Amanda Hui Qi
Wilm, Andreas
Nagarajan, Niranjan
author_facet Li, Chenhao
Chng, Kern Rei
Boey, Esther Jia Hui
Ng, Amanda Hui Qi
Wilm, Andreas
Nagarajan, Niranjan
author_sort Li, Chenhao
collection PubMed
description BACKGROUND: Nanopore sequencing provides a rapid, cheap and portable real-time sequencing platform with the potential to revolutionize genomics. However, several applications are limited by relatively high single-read error rates (>10 %), including RNA-seq, haplotype sequencing and 16S sequencing. RESULTS: We developed the Intramolecular-ligated Nanopore Consensus Sequencing (INC-Seq) as a strategy for obtaining long and accurate nanopore reads, starting with low input DNA. Applying INC-Seq for 16S rRNA-based bacterial profiling generated full-length amplicon sequences with a median accuracy >97 %. CONCLUSIONS: INC-Seq reads enabled accurate species-level classification, identification of species at 0.1 % abundance and robust quantification of relative abundances, providing a cheap and effective approach for pathogen detection and microbiome profiling on the MinION system. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13742-016-0140-7) contains supplementary material, which is available to authorized users.
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spelling pubmed-49702892016-08-03 INC-Seq: accurate single molecule reads using nanopore sequencing Li, Chenhao Chng, Kern Rei Boey, Esther Jia Hui Ng, Amanda Hui Qi Wilm, Andreas Nagarajan, Niranjan Gigascience Methodology BACKGROUND: Nanopore sequencing provides a rapid, cheap and portable real-time sequencing platform with the potential to revolutionize genomics. However, several applications are limited by relatively high single-read error rates (>10 %), including RNA-seq, haplotype sequencing and 16S sequencing. RESULTS: We developed the Intramolecular-ligated Nanopore Consensus Sequencing (INC-Seq) as a strategy for obtaining long and accurate nanopore reads, starting with low input DNA. Applying INC-Seq for 16S rRNA-based bacterial profiling generated full-length amplicon sequences with a median accuracy >97 %. CONCLUSIONS: INC-Seq reads enabled accurate species-level classification, identification of species at 0.1 % abundance and robust quantification of relative abundances, providing a cheap and effective approach for pathogen detection and microbiome profiling on the MinION system. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13742-016-0140-7) contains supplementary material, which is available to authorized users. BioMed Central 2016-08-02 /pmc/articles/PMC4970289/ /pubmed/27485345 http://dx.doi.org/10.1186/s13742-016-0140-7 Text en © The Author(s). 2016 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
Li, Chenhao
Chng, Kern Rei
Boey, Esther Jia Hui
Ng, Amanda Hui Qi
Wilm, Andreas
Nagarajan, Niranjan
INC-Seq: accurate single molecule reads using nanopore sequencing
title INC-Seq: accurate single molecule reads using nanopore sequencing
title_full INC-Seq: accurate single molecule reads using nanopore sequencing
title_fullStr INC-Seq: accurate single molecule reads using nanopore sequencing
title_full_unstemmed INC-Seq: accurate single molecule reads using nanopore sequencing
title_short INC-Seq: accurate single molecule reads using nanopore sequencing
title_sort inc-seq: accurate single molecule reads using nanopore sequencing
topic Methodology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4970289/
https://www.ncbi.nlm.nih.gov/pubmed/27485345
http://dx.doi.org/10.1186/s13742-016-0140-7
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