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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...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2016
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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. |
format | Online Article Text |
id | pubmed-4970289 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
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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