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. 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

Affiliations

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

Benjamin Istace et al. Gigascience. .

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.

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Figures

Figure 1:
Figure 1:
Identity distribution of Nanopore reads. Percent identity of the aligned MinION 1D (red bars) and 2D (green bars) reads. The MinION reads were aligned using LAST software. (a) R7.3 chemistry. (b) R9 chemistry.
Figure 2:
Figure 2:
Feature composition of the S288C assemblies, assembly and quality metrics, and assembler running statistics. The feature content of the best S288C assemblies for each assembler is shown in the left part of the figure. The feature composition was obtained by aligning each assembly to the S288C reference genome. Assembly and quality metrics for each assembly, obtained by using Quast, are shown in the middle part of the figure. The running time and the memory usage of each assembler are shown in the right part of the figure.
Figure 3:
Figure 3:
Cartography of the Ty transposon family. First and second tracks show, respectively, the percentage identity of the SMARTdenovo S288C assembly before and after polishing with Illumina paired-end reads using Pilon. The third track shows the 80th percentile number of contigs obtained for each strain and for all chromosomes. The remaining tracks show the density of Ty transposons or positions of the Ty1, Ty2, Ty3, Ty4, and Ty5 transposons across all the yeast strains. The red dot on the karyotype track shows the position of the rDNA cluster.

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