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Benchmarking of long-read correction methods

Third-generation sequencing technologies provided by Pacific Biosciences and Oxford Nanopore Technologies generate read lengths in the scale of kilobasepairs. However, these reads display high error rates, and correction steps are necessary to realize their great potential in genomics and transcript...

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Autores principales: Dohm, Juliane C, Peters, Philipp, Stralis-Pavese, Nancy, Himmelbauer, Heinz
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7671305/
https://www.ncbi.nlm.nih.gov/pubmed/33575591
http://dx.doi.org/10.1093/nargab/lqaa037
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author Dohm, Juliane C
Peters, Philipp
Stralis-Pavese, Nancy
Himmelbauer, Heinz
author_facet Dohm, Juliane C
Peters, Philipp
Stralis-Pavese, Nancy
Himmelbauer, Heinz
author_sort Dohm, Juliane C
collection PubMed
description Third-generation sequencing technologies provided by Pacific Biosciences and Oxford Nanopore Technologies generate read lengths in the scale of kilobasepairs. However, these reads display high error rates, and correction steps are necessary to realize their great potential in genomics and transcriptomics. Here, we compare properties of PacBio and Nanopore data and assess correction methods by Canu, MARVEL and proovread in various combinations. We found total error rates of around 13% in the raw datasets. PacBio reads showed a high rate of insertions (around 8%) whereas Nanopore reads showed similar rates for substitutions, insertions and deletions of around 4% each. In data from both technologies the errors were uniformly distributed along reads apart from noisy 5′ ends, and homopolymers appeared among the most over-represented kmers relative to a reference. Consensus correction using read overlaps reduced error rates to about 1% when using Canu or MARVEL after patching. The lowest error rate in Nanopore data (0.45%) was achieved by applying proovread on MARVEL-patched data including Illumina short-reads, and the lowest error rate in PacBio data (0.42%) was the result of Canu correction with minimap2 alignment after patching. Our study provides valuable insights and benchmarks regarding long-read data and correction methods.
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spelling pubmed-76713052021-02-10 Benchmarking of long-read correction methods Dohm, Juliane C Peters, Philipp Stralis-Pavese, Nancy Himmelbauer, Heinz NAR Genom Bioinform Standard Article Third-generation sequencing technologies provided by Pacific Biosciences and Oxford Nanopore Technologies generate read lengths in the scale of kilobasepairs. However, these reads display high error rates, and correction steps are necessary to realize their great potential in genomics and transcriptomics. Here, we compare properties of PacBio and Nanopore data and assess correction methods by Canu, MARVEL and proovread in various combinations. We found total error rates of around 13% in the raw datasets. PacBio reads showed a high rate of insertions (around 8%) whereas Nanopore reads showed similar rates for substitutions, insertions and deletions of around 4% each. In data from both technologies the errors were uniformly distributed along reads apart from noisy 5′ ends, and homopolymers appeared among the most over-represented kmers relative to a reference. Consensus correction using read overlaps reduced error rates to about 1% when using Canu or MARVEL after patching. The lowest error rate in Nanopore data (0.45%) was achieved by applying proovread on MARVEL-patched data including Illumina short-reads, and the lowest error rate in PacBio data (0.42%) was the result of Canu correction with minimap2 alignment after patching. Our study provides valuable insights and benchmarks regarding long-read data and correction methods. Oxford University Press 2020-05-25 /pmc/articles/PMC7671305/ /pubmed/33575591 http://dx.doi.org/10.1093/nargab/lqaa037 Text en © The Author(s) 2019. Published by Oxford University Press on behalf of NAR Genomics and Bioinformatics. http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Standard Article
Dohm, Juliane C
Peters, Philipp
Stralis-Pavese, Nancy
Himmelbauer, Heinz
Benchmarking of long-read correction methods
title Benchmarking of long-read correction methods
title_full Benchmarking of long-read correction methods
title_fullStr Benchmarking of long-read correction methods
title_full_unstemmed Benchmarking of long-read correction methods
title_short Benchmarking of long-read correction methods
title_sort benchmarking of long-read correction methods
topic Standard Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7671305/
https://www.ncbi.nlm.nih.gov/pubmed/33575591
http://dx.doi.org/10.1093/nargab/lqaa037
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