Assembly methods for nanopore-based metagenomic sequencing: a comparative study

Abstract Metagenomic sequencing has allowed for the recovery of previously unexplored microbial genomes. Whereas short-read sequencing platforms often result in highly fragmented metagenomes, nanopore-based sequencers could lead to more contiguous assemblies due to their potential to generate long r...

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Main Authors: Adriel Latorre-Pérez, Pascual Villalba-Bermell, Javier Pascual, Cristina Vilanova
Format: Article
Language:English
Published: Nature Publishing Group 2020-08-01
Series:Scientific Reports
Online Access:https://doi.org/10.1038/s41598-020-70491-3
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spelling doaj-ed3be38cf7654157b93e181259da1bfc2021-08-15T11:19:13ZengNature Publishing GroupScientific Reports2045-23222020-08-0110111410.1038/s41598-020-70491-3Assembly methods for nanopore-based metagenomic sequencing: a comparative studyAdriel Latorre-Pérez0Pascual Villalba-Bermell1Javier Pascual2Cristina Vilanova3Darwin Bioprospecting Excellence S.L.Darwin Bioprospecting Excellence S.L.Darwin Bioprospecting Excellence S.L.Darwin Bioprospecting Excellence S.L.Abstract Metagenomic sequencing has allowed for the recovery of previously unexplored microbial genomes. Whereas short-read sequencing platforms often result in highly fragmented metagenomes, nanopore-based sequencers could lead to more contiguous assemblies due to their potential to generate long reads. Nevertheless, there is a lack of updated and systematic studies evaluating the performance of different assembly tools on nanopore data. In this study, we have benchmarked the ability of different assemblers to reconstruct two different commercially-available mock communities that have been sequenced using Oxford Nanopore Technologies platforms. Among the tested tools, only metaFlye, Raven, and Canu performed well in all the datasets. These tools retrieved highly contiguous genomes (or even complete genomes) directly from the metagenomic data. Despite the intrinsic high error of nanopore sequencing, final assemblies reached high accuracy (~ 99.5 to 99.8% of consensus accuracy). Polishing strategies demonstrated to be necessary for reducing the number of indels, and this had an impact on the prediction of biosynthetic gene clusters. Correction with high quality short reads did not always result in higher quality draft assemblies. Overall, nanopore metagenomic sequencing data-adapted to MinION’s current output-proved sufficient for assembling and characterizing low-complexity microbial communities.https://doi.org/10.1038/s41598-020-70491-3
collection DOAJ
language English
format Article
sources DOAJ
author Adriel Latorre-Pérez
Pascual Villalba-Bermell
Javier Pascual
Cristina Vilanova
spellingShingle Adriel Latorre-Pérez
Pascual Villalba-Bermell
Javier Pascual
Cristina Vilanova
Assembly methods for nanopore-based metagenomic sequencing: a comparative study
Scientific Reports
author_facet Adriel Latorre-Pérez
Pascual Villalba-Bermell
Javier Pascual
Cristina Vilanova
author_sort Adriel Latorre-Pérez
title Assembly methods for nanopore-based metagenomic sequencing: a comparative study
title_short Assembly methods for nanopore-based metagenomic sequencing: a comparative study
title_full Assembly methods for nanopore-based metagenomic sequencing: a comparative study
title_fullStr Assembly methods for nanopore-based metagenomic sequencing: a comparative study
title_full_unstemmed Assembly methods for nanopore-based metagenomic sequencing: a comparative study
title_sort assembly methods for nanopore-based metagenomic sequencing: a comparative study
publisher Nature Publishing Group
series Scientific Reports
issn 2045-2322
publishDate 2020-08-01
description Abstract Metagenomic sequencing has allowed for the recovery of previously unexplored microbial genomes. Whereas short-read sequencing platforms often result in highly fragmented metagenomes, nanopore-based sequencers could lead to more contiguous assemblies due to their potential to generate long reads. Nevertheless, there is a lack of updated and systematic studies evaluating the performance of different assembly tools on nanopore data. In this study, we have benchmarked the ability of different assemblers to reconstruct two different commercially-available mock communities that have been sequenced using Oxford Nanopore Technologies platforms. Among the tested tools, only metaFlye, Raven, and Canu performed well in all the datasets. These tools retrieved highly contiguous genomes (or even complete genomes) directly from the metagenomic data. Despite the intrinsic high error of nanopore sequencing, final assemblies reached high accuracy (~ 99.5 to 99.8% of consensus accuracy). Polishing strategies demonstrated to be necessary for reducing the number of indels, and this had an impact on the prediction of biosynthetic gene clusters. Correction with high quality short reads did not always result in higher quality draft assemblies. Overall, nanopore metagenomic sequencing data-adapted to MinION’s current output-proved sufficient for assembling and characterizing low-complexity microbial communities.
url https://doi.org/10.1038/s41598-020-70491-3
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