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Title: Systematic and stochastic influences on the performance of the MinION nanopore sequencer across a range of nucleotide bias

Journal Article · · Scientific Reports

Emerging sequencing technologies are allowing us to characterize environmental, clinical and laboratory samples with increasing speed and detail, including real-time analysis and interpretation of data. One example of this is being able to rapidly and accurately detect a wide range of pathogenic organisms, both in the clinic and the field. Genomes can have radically different GC content however, such that accurate sequence analysis can be challenging depending upon the technology used. Here, we have characterized the performance of the Oxford MinION nanopore sequencer for detection and evaluation of organisms with a range of genomic nucleotide bias. We have diagnosed the quality of base-calling across individual reads and discovered that the position within the read affects base-calling and quality scores. Finally, we have evaluated the performance of the current state-of-the-art neural network-based MinION basecaller, characterizing its behavior with respect to systemic errors as well as context- and sequence-specific errors. Overall, we present a detailed characterization the capabilities of the MinION in terms of generating high-accuracy sequence data from genomes with a wide range of nucleotide content. This study provides a framework for designing the appropriate experiments that are the likely to lead to accurate and rapid field-forward diagnostics.

Research Organization:
Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)
Sponsoring Organization:
USDOE National Nuclear Security Administration (NNSA)
Grant/Contract Number:
AC04-94AL85000
OSTI ID:
1430904
Report Number(s):
SAND-2018-0811J; 661556
Journal Information:
Scientific Reports, Vol. 8, Issue 1; ISSN 2045-2322
Publisher:
Nature Publishing GroupCopyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 36 works
Citation information provided by
Web of Science

References (34)

Genome assembly using Nanopore-guided long and error-free DNA reads journal April 2015
Rapid metagenomic identification of viral pathogens in clinical samples by real-time nanopore sequencing analysis journal September 2015
The Oxford Nanopore MinION: delivery of nanopore sequencing to the genomics community journal November 2016
Oxford Nanopore sequencing, hybrid error correction, and de novo assembly of a eukaryotic genome journal October 2015
Rapid draft sequencing and real-time nanopore sequencing in a hospital outbreak of Salmonella journal May 2015
Improved data analysis for the MinION nanopore sequencer journal February 2015
Nanopore sequencing enables near-complete de novo assembly of Saccharomyces cerevisiae reference strain CEN.PK113-7D journal September 2017
Real-Time DNA Sequencing from Single Polymerase Molecules journal January 2009
Real-time, portable genome sequencing for Ebola surveillance journal February 2016
Summarizing and correcting the GC content bias in high-throughput sequencing journal February 2012
De Novo Assembly of a New Solanum pennellii Accession Using Nanopore Sequencing journal October 2017
BEDTools: a flexible suite of utilities for comparing genomic features journal January 2010
Effects of GC Bias in Next-Generation-Sequencing Data on De Novo Genome Assembly journal April 2013
MinION nanopore sequencing of an influenza genome journal January 2015
Multiplex PCR method for MinION and Illumina sequencing of Zika and other virus genomes directly from clinical samples journal May 2017
Analyzing and minimizing PCR amplification bias in Illumina sequencing libraries journal January 2011
Characterizing and measuring bias in sequence data journal January 2013
Rapid de novo assembly of the European eel genome from nanopore sequencing reads journal August 2017
PacBio Sequencing and Its Applications journal October 2015
Comprehensive transcriptome analysis using synthetic long-read sequencing reveals molecular co-association of distant splicing events journal May 2015
Assembling large genomes with single-molecule sequencing and locality-sensitive hashing journal May 2015
High-Quality de Novo Genome Assembly of the Dekkera bruxellensis Yeast Using Nanopore MinION Sequencing journal August 2017
Fast and accurate long-read alignment with Burrows–Wheeler transform journal January 2010
Real-time selective sequencing using nanopore technology journal July 2016
Mapping DNA methylation with high-throughput nanopore sequencing journal February 2017
Benchmarking of the Oxford Nanopore MinION sequencing for quantitative and qualitative assessment of cDNA populations journal August 2016
Scaffolding of a bacterial genome using MinION nanopore sequencing journal July 2015
The Sequence Alignment/Map format and SAMtools journal June 2009
Assessing the performance of the Oxford Nanopore Technologies MinION journal March 2015
Direct detection of DNA methylation during single-molecule, real-time sequencing journal May 2010
Nanopore sequencing and assembly of a human genome with ultra-long reads journal January 2018
Production of viable chicken by allogeneic transplantation of primordial germ cells induced from somatic cells journal May 2021
Same-Day Diagnostic and Surveillance Data for Tuberculosis via Whole-Genome Sequencing of Direct Respiratory Samples journal March 2017
An Improved Filtering Algorithm for Big Read Datasets preprint January 2016

Cited By (12)

Increasing the accuracy of nanopore DNA sequencing using a time-varying cross membrane voltage journal April 2019
Long reads: their purpose and place journal May 2018
Ultraplexing: increasing the efficiency of long-read sequencing for hybrid assembly with k-mer-based multiplexing journal March 2020
Long-Read Sequencing Emerging in Medical Genetics journal May 2019
Human Herpesvirus Sequencing in the Genomic Era: The Growing Ranks of the Herpetic Legion journal October 2019
Long reads: their purpose and place. text January 2018
Human Herpesvirus Sequencing in the Genomic Era: The Growing Ranks of the Herpetic Legion text January 2019
Real-Time Selective Sequencing with RUBRIC: Read Until with Basecall and Reference-Informed Criteria journal August 2019
De Novo Clustering of Long-Read Transcriptome Data Using a Greedy, Quality Value-Based Algorithm journal April 2020
High-Fidelity Nanopore Sequencing of Ultra-Short DNA Targets journal April 2019
More accurate nanopore sequencing journal May 2019
Assessment of low-coverage nanopore long read sequencing for SNP genotyping in doubled haploid canola (Brassica napus L.) journal June 2019

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