R2C2+UMI: Combining concatemeric consensus sequencing with unique molecular identifiers enables ultra-accurate sequencing of amplicons on Oxford Nanopore Technologies sequencers Article Swipe
Dori Z.Q. Deng
,
Jack Verhage
,
Celine Neudorf
,
Russell Corbett‐Detig
,
Honey Mekonen
,
Peter J. Castaldi
,
Christopher Vollmers
·
YOU?
·
· 2023
· Open Access
·
· DOI: https://doi.org/10.1101/2023.08.19.553937
YOU?
·
· 2023
· Open Access
·
· DOI: https://doi.org/10.1101/2023.08.19.553937
The sequencing of PCR amplicons is a core application of high-throughput sequencing technology. Using unique molecular identifiers (UMIs), individual amplified molecules can be sequenced to very high accuracy on an Illumina sequencer. However, Illumina sequencers have limited read length and are therefore restricted to sequencing amplicons shorter than 600bp unless using inefficient synthetic long-read approaches. Native long-read sequencers from Pacific Biosciences and Oxford Nanopore Technologies can, using consensus read approaches, match or exceed Illumina quality while achieving much longer read lengths. Using a circularization-based concatemeric consensus sequencing approach (R2C2) paired with UMIs (R2C2+UMI) we show that we can sequence ∼550nt antibody heavy-chain (IGH) and ∼1500nt 16S amplicons at accuracies up to and exceeding Q50 (<1 error in 100,0000 sequenced bases), which exceeds accuracies of UMI-supported Illumina paired sequencing as well as synthetic long-read approaches.
Related Topics
Concepts
Nanopore sequencing
Amplicon
Identifier
Computational biology
Biology
Computer science
DNA sequencing
Genetics
Computer network
Gene
Polymerase chain reaction
Metadata
- Type
- preprint
- Language
- en
- Landing Page
- https://doi.org/10.1101/2023.08.19.553937
- https://www.biorxiv.org/content/biorxiv/early/2023/08/21/2023.08.19.553937.full.pdf
- OA Status
- green
- Cited By
- 6
- References
- 35
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W4386030837
All OpenAlex metadata
Raw OpenAlex JSON
- OpenAlex ID
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- DOI
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https://doi.org/10.1101/2023.08.19.553937Digital Object Identifier
- Title
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R2C2+UMI: Combining concatemeric consensus sequencing with unique molecular identifiers enables ultra-accurate sequencing of amplicons on Oxford Nanopore Technologies sequencersWork title
- Type
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preprintOpenAlex work type
- Language
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enPrimary language
- Publication year
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2023Year of publication
- Publication date
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2023-08-21Full publication date if available
- Authors
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Dori Z.Q. Deng, Jack Verhage, Celine Neudorf, Russell Corbett‐Detig, Honey Mekonen, Peter J. Castaldi, Christopher VollmersList of authors in order
- Landing page
-
https://doi.org/10.1101/2023.08.19.553937Publisher landing page
- PDF URL
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https://www.biorxiv.org/content/biorxiv/early/2023/08/21/2023.08.19.553937.full.pdfDirect link to full text PDF
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YesWhether a free full text is available
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greenOpen access status per OpenAlex
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https://www.biorxiv.org/content/biorxiv/early/2023/08/21/2023.08.19.553937.full.pdfDirect OA link when available
- Concepts
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Nanopore sequencing, Amplicon, Identifier, Computational biology, Biology, Computer science, DNA sequencing, Genetics, Computer network, Gene, Polymerase chain reactionTop concepts (fields/topics) attached by OpenAlex
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6Total citation count in OpenAlex
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2025: 1, 2024: 5Per-year citation counts (last 5 years)
- References (count)
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35Number of works referenced by this work
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-
10Other works algorithmically related by OpenAlex
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