Systematic and stochastic influences on the performance of the MinION nanopore sequencer across a range of nucleotide bias
Abstract
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.
- Authors:
-
- Sandia National Lab. (SNL-CA), Livermore, CA (United States)
- Publication Date:
- Research Org.:
- Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)
- Sponsoring Org.:
- USDOE National Nuclear Security Administration (NNSA)
- OSTI Identifier:
- 1430904
- Report Number(s):
- SAND-2018-0811J
Journal ID: ISSN 2045-2322; 661556
- Grant/Contract Number:
- AC04-94AL85000
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Scientific Reports
- Additional Journal Information:
- Journal Volume: 8; Journal Issue: 1; Journal ID: ISSN 2045-2322
- Publisher:
- Nature Publishing Group
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES
Citation Formats
Krishnakumar, Raga, Sinha, Anupama, Bird, Sara W., Jayamohan, Harikrishnan, Edwards, Harrison S., Schoeniger, Joseph S., Patel, Kamlesh D., Branda, Steven S., and Bartsch, Michael S. Systematic and stochastic influences on the performance of the MinION nanopore sequencer across a range of nucleotide bias. United States: N. p., 2018.
Web. doi:10.1038/s41598-018-21484-w.
Krishnakumar, Raga, Sinha, Anupama, Bird, Sara W., Jayamohan, Harikrishnan, Edwards, Harrison S., Schoeniger, Joseph S., Patel, Kamlesh D., Branda, Steven S., & Bartsch, Michael S. Systematic and stochastic influences on the performance of the MinION nanopore sequencer across a range of nucleotide bias. United States. https://doi.org/10.1038/s41598-018-21484-w
Krishnakumar, Raga, Sinha, Anupama, Bird, Sara W., Jayamohan, Harikrishnan, Edwards, Harrison S., Schoeniger, Joseph S., Patel, Kamlesh D., Branda, Steven S., and Bartsch, Michael S. Fri .
"Systematic and stochastic influences on the performance of the MinION nanopore sequencer across a range of nucleotide bias". United States. https://doi.org/10.1038/s41598-018-21484-w. https://www.osti.gov/servlets/purl/1430904.
@article{osti_1430904,
title = {Systematic and stochastic influences on the performance of the MinION nanopore sequencer across a range of nucleotide bias},
author = {Krishnakumar, Raga and Sinha, Anupama and Bird, Sara W. and Jayamohan, Harikrishnan and Edwards, Harrison S. and Schoeniger, Joseph S. and Patel, Kamlesh D. and Branda, Steven S. and Bartsch, Michael S.},
abstractNote = {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.},
doi = {10.1038/s41598-018-21484-w},
journal = {Scientific Reports},
number = 1,
volume = 8,
place = {United States},
year = {Fri Feb 16 00:00:00 EST 2018},
month = {Fri Feb 16 00:00:00 EST 2018}
}
Web of Science
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