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Assembly-free single-molecule nanopore sequencing recovers complete virus genomes from natural microbial communities

View ORCID ProfileJohn Beaulaurier, Elaine Luo, John Eppley, Paul Den Uyl, Xiaoguang Dai, Daniel J Turner, Matthew Pendelton, Sissel Juul, Eoghan Harrington, Edward F. DeLong
doi: https://doi.org/10.1101/619684
John Beaulaurier
1Oxford Nanopore Technologies Inc., San Francisco, CA, USA
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  • ORCID record for John Beaulaurier
Elaine Luo
2Daniel K. Inouye Center for Microbial Oceanography: Research and Education, University of Hawaii, Honolulu, Hawaii, USA
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John Eppley
2Daniel K. Inouye Center for Microbial Oceanography: Research and Education, University of Hawaii, Honolulu, Hawaii, USA
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Paul Den Uyl
2Daniel K. Inouye Center for Microbial Oceanography: Research and Education, University of Hawaii, Honolulu, Hawaii, USA
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Xiaoguang Dai
3Oxford Nanopore Technologies Inc., New York, New York, USA
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Daniel J Turner
4Oxford Nanopore Technologies Ltd., Oxford, UK
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Matthew Pendelton
3Oxford Nanopore Technologies Inc., New York, New York, USA
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Sissel Juul
3Oxford Nanopore Technologies Inc., New York, New York, USA
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Eoghan Harrington
3Oxford Nanopore Technologies Inc., New York, New York, USA
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Edward F. DeLong
2Daniel K. Inouye Center for Microbial Oceanography: Research and Education, University of Hawaii, Honolulu, Hawaii, USA
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  • For correspondence: edelong{at}hawaii.edu
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Abstract

Viruses are the most abundant biological entities on Earth, and play key roles in host ecology, evolution, and horizontal gene transfer. Despite recent progress in viral metagenomics, the inherent genetic complexity of virus populations still poses technical difficulties for recovering complete virus genomes from natural assemblages. To address these challenges, we developed an assembly-free, single-molecule nanopore sequencing approach enabling direct recovery of high-quality viral genome sequences from environmental samples. Our method yielded over a thousand high quality, full-length draft virus genome sequences that could not be fully recovered using short read assembly approaches applied to the same samples. Additionally, novel DNA sequences were discovered whose repeat structures, gene contents and concatemer lengths suggested that they represent phage-inducible chromosomal islands that were packaged as concatemers within phage particles. Our new approach provided novel insight into genome structures, population biology, and ecology of naturally occurring viruses and viral parasites.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted April 26, 2019.
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Assembly-free single-molecule nanopore sequencing recovers complete virus genomes from natural microbial communities
John Beaulaurier, Elaine Luo, John Eppley, Paul Den Uyl, Xiaoguang Dai, Daniel J Turner, Matthew Pendelton, Sissel Juul, Eoghan Harrington, Edward F. DeLong
bioRxiv 619684; doi: https://doi.org/10.1101/619684
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Assembly-free single-molecule nanopore sequencing recovers complete virus genomes from natural microbial communities
John Beaulaurier, Elaine Luo, John Eppley, Paul Den Uyl, Xiaoguang Dai, Daniel J Turner, Matthew Pendelton, Sissel Juul, Eoghan Harrington, Edward F. DeLong
bioRxiv 619684; doi: https://doi.org/10.1101/619684

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