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Gigascience. 2017 Feb 1;6(2):1-13. doi: 10.1093/gigascience/giw018.

de novo assembly and population genomic survey of natural yeast isolates with the Oxford Nanopore MinION sequencer.

Author information

1
Commissariat à l'Energie Atomique et aux Energies Alternatives (CEA), Institut de Génomique (IG), Genoscope, BP5706, 91057 Evry, France.
2
Université de Strasbourg, CNRS, GMGM UMR 7156, F-67000 Strasbourg, France.
3
Institute of Research on Cancer and Ageing of Nice (IRCAN), CNRS UMR 7284-INSERM U1081, Faculté de Médecine, Université de Nice Sophia Antipolis, Nice, France.
4
Université d'Evry Val d'Essonne, UMR 8030, CP5706, 91057 Evry, France.
5
Centre National de Recherche Scientifique (CNRS), UMR 8030, CP5706, 91057 Evry, France.

Abstract

BACKGROUND:

Oxford Nanopore Technologies Ltd (Oxford, UK) have recently commercialized MinION, a small single-molecule nanopore sequencer, that offers the possibility of sequencing long DNA fragments from small genomes in a matter of seconds. The Oxford Nanopore technology is truly disruptive; it has the potential to revolutionize genomic applications due to its portability, low cost, and ease of use compared with existing long reads sequencing technologies. The MinION sequencer enables the rapid sequencing of small eukaryotic genomes, such as the yeast genome. Combined with existing assembler algorithms, near complete genome assemblies can be generated and comprehensive population genomic analyses can be performed.

RESULTS:

Here, we resequenced the genome of the Saccharomyces cerevisiae S288C strain to evaluate the performance of nanopore-only assemblers. Then we de novo sequenced and assembled the genomes of 21 isolates representative of the S. cerevisiae genetic diversity using the MinION platform. The contiguity of our assemblies was 14 times higher than the Illumina-only assemblies and we obtained one or two long contigs for 65 % of the chromosomes. This high contiguity allowed us to accurately detect large structural variations across the 21 studied genomes.

CONCLUSION:

Because of the high completeness of the nanopore assemblies, we were able to produce a complete cartography of transposable elements insertions and inspect structural variants that are generally missed using a short-read sequencing strategy. Our analyses show that the Oxford Nanopore technology is already usable for de novo sequencing and assembly; however, non-random errors in homopolymers require polishing the consensus using an alternate sequencing technology.

KEYWORDS:

Genome finishing; MinION device; Nanopore sequencing; Oxford Nanopore; Structural variations; Transposable elements; de novo assembly

PMID:
28369459
PMCID:
PMC5466710
DOI:
10.1093/gigascience/giw018
[Indexed for MEDLINE]
Free PMC Article

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