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De novo assembly of middle-sized genome using MinION and Illumina sequencers

BACKGROUND: The plastid acquisition by secondary endosymbiosis is a driving force for the algal evolution, and the comparative genomics was required to examine the genomic change of symbiont. Therefore, we established a pipeline of a de novo assembly of middle-sized genomes at a low cost and with hi...

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Autores principales: Minei, Ryuhei, Hoshina, Ryo, Ogura, Atsushi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6154909/
https://www.ncbi.nlm.nih.gov/pubmed/30249180
http://dx.doi.org/10.1186/s12864-018-5067-1
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author Minei, Ryuhei
Hoshina, Ryo
Ogura, Atsushi
author_facet Minei, Ryuhei
Hoshina, Ryo
Ogura, Atsushi
author_sort Minei, Ryuhei
collection PubMed
description BACKGROUND: The plastid acquisition by secondary endosymbiosis is a driving force for the algal evolution, and the comparative genomics was required to examine the genomic change of symbiont. Therefore, we established a pipeline of a de novo assembly of middle-sized genomes at a low cost and with high quality using long and short reads. RESULTS: We sequenced symbiotic algae Chlorella variabilis using Oxfofrd Nanopore MinION as the long-read sequencer and Illumina HiSeq 4000 as the short-read sequencer and then assembled the genomes under various conditions. Subsequently, we evaluated these assemblies by the gene model quality and RNA-seq mapping rate. We found that long-read only assembly could not be suitable for the comparative genomics studies, but with short reads, we could obtain the acceptable assembly. On the basis of this result, we established the pipeline of de novo assembly for middle-sized algal genome using MinION. CONCLUSIONS: The genomic change during the early stages of plastid acquisition can now be revealed by sequencing and comparing many algal genomes. Moreover, this pipeline offers a solution for the assembly of various middle-sized eukaryotic genomes with high-quality and ease. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12864-018-5067-1) contains supplementary material, which is available to authorized users.
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spelling pubmed-61549092018-09-26 De novo assembly of middle-sized genome using MinION and Illumina sequencers Minei, Ryuhei Hoshina, Ryo Ogura, Atsushi BMC Genomics Methodology Article BACKGROUND: The plastid acquisition by secondary endosymbiosis is a driving force for the algal evolution, and the comparative genomics was required to examine the genomic change of symbiont. Therefore, we established a pipeline of a de novo assembly of middle-sized genomes at a low cost and with high quality using long and short reads. RESULTS: We sequenced symbiotic algae Chlorella variabilis using Oxfofrd Nanopore MinION as the long-read sequencer and Illumina HiSeq 4000 as the short-read sequencer and then assembled the genomes under various conditions. Subsequently, we evaluated these assemblies by the gene model quality and RNA-seq mapping rate. We found that long-read only assembly could not be suitable for the comparative genomics studies, but with short reads, we could obtain the acceptable assembly. On the basis of this result, we established the pipeline of de novo assembly for middle-sized algal genome using MinION. CONCLUSIONS: The genomic change during the early stages of plastid acquisition can now be revealed by sequencing and comparing many algal genomes. Moreover, this pipeline offers a solution for the assembly of various middle-sized eukaryotic genomes with high-quality and ease. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12864-018-5067-1) contains supplementary material, which is available to authorized users. BioMed Central 2018-09-24 /pmc/articles/PMC6154909/ /pubmed/30249180 http://dx.doi.org/10.1186/s12864-018-5067-1 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
Minei, Ryuhei
Hoshina, Ryo
Ogura, Atsushi
De novo assembly of middle-sized genome using MinION and Illumina sequencers
title De novo assembly of middle-sized genome using MinION and Illumina sequencers
title_full De novo assembly of middle-sized genome using MinION and Illumina sequencers
title_fullStr De novo assembly of middle-sized genome using MinION and Illumina sequencers
title_full_unstemmed De novo assembly of middle-sized genome using MinION and Illumina sequencers
title_short De novo assembly of middle-sized genome using MinION and Illumina sequencers
title_sort de novo assembly of middle-sized genome using minion and illumina sequencers
topic Methodology Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6154909/
https://www.ncbi.nlm.nih.gov/pubmed/30249180
http://dx.doi.org/10.1186/s12864-018-5067-1
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