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Highly parallel direct RNA sequencing on an array of nanopores

Daniel R. Garalde, Elizabeth A. Snell, Daniel Jachimowicz, Andrew J. Heron, Mark Bruce, Joseph Lloyd, Anthony Warland, Nadia Pantic, Tigist Admassu, Jonah Ciccone, Sabrina Serra, Jemma Keenan, Samuel Martin, Luke McNeill, Jayne Wallace, Lakmal Jayasinghe, Chris Wright, Javier Blasco, Botond Sipos, Stephen Young, Sissel Juul, James Clarke, Daniel J Turner
doi: https://doi.org/10.1101/068809
Daniel R. Garalde
1Oxford Nanopore Technologies Ltd., Oxford, UK
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Elizabeth A. Snell
1Oxford Nanopore Technologies Ltd., Oxford, UK
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Daniel Jachimowicz
1Oxford Nanopore Technologies Ltd., Oxford, UK
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Andrew J. Heron
1Oxford Nanopore Technologies Ltd., Oxford, UK
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Mark Bruce
1Oxford Nanopore Technologies Ltd., Oxford, UK
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Joseph Lloyd
1Oxford Nanopore Technologies Ltd., Oxford, UK
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Anthony Warland
1Oxford Nanopore Technologies Ltd., Oxford, UK
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Nadia Pantic
1Oxford Nanopore Technologies Ltd., Oxford, UK
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Tigist Admassu
1Oxford Nanopore Technologies Ltd., Oxford, UK
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Jonah Ciccone
1Oxford Nanopore Technologies Ltd., Oxford, UK
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Sabrina Serra
1Oxford Nanopore Technologies Ltd., Oxford, UK
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Jemma Keenan
1Oxford Nanopore Technologies Ltd., Oxford, UK
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Samuel Martin
1Oxford Nanopore Technologies Ltd., Oxford, UK
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Luke McNeill
1Oxford Nanopore Technologies Ltd., Oxford, UK
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Jayne Wallace
1Oxford Nanopore Technologies Ltd., Oxford, UK
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Lakmal Jayasinghe
1Oxford Nanopore Technologies Ltd., Oxford, UK
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Chris Wright
1Oxford Nanopore Technologies Ltd., Oxford, UK
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Javier Blasco
1Oxford Nanopore Technologies Ltd., Oxford, UK
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Botond Sipos
1Oxford Nanopore Technologies Ltd., Oxford, UK
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Stephen Young
1Oxford Nanopore Technologies Ltd., Oxford, UK
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Sissel Juul
2Oxford Nanopore Technologies Inc., New York, USA
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James Clarke
1Oxford Nanopore Technologies Ltd., Oxford, UK
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Daniel J Turner
1Oxford Nanopore Technologies Ltd., Oxford, UK
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  • For correspondence: daniel.turner@nanoporetech.com
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Abstract

Ribonucleic acid sequencing can allow us to monitor the RNAs present in a sample. This enables us to detect the presence and nucleotide sequence of viruses, or to build a picture of how active transcriptional processes are changing – information that is useful for understanding the status and function of a sample. Oxford Nanopore Technologies’ sequencing technology is capable of electronically analysing a sample’s DNA directly, and in real-time. In this manuscript we demonstrate the ability of an array of nanopores to sequence RNA directly, and we apply it to a range of biological situations. Nanopore technology is the only available sequencing technology that can sequence RNA directly, rather than depending on reverse transcription and PCR. There are several potential advantages of this approach over other RNA-seq strategies, including the absence of amplification and reverse transcription biases, the ability to detect nucleotide analogues and the ability to generate full-length, strand-specific RNA sequences. Direct RNA sequencing is a completely new way of analysing the sequence of RNA samples and it will improve the ease and speed of RNA analysis, while yielding richer biological information.

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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 August 12, 2016.
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Highly parallel direct RNA sequencing on an array of nanopores
Daniel R. Garalde, Elizabeth A. Snell, Daniel Jachimowicz, Andrew J. Heron, Mark Bruce, Joseph Lloyd, Anthony Warland, Nadia Pantic, Tigist Admassu, Jonah Ciccone, Sabrina Serra, Jemma Keenan, Samuel Martin, Luke McNeill, Jayne Wallace, Lakmal Jayasinghe, Chris Wright, Javier Blasco, Botond Sipos, Stephen Young, Sissel Juul, James Clarke, Daniel J Turner
bioRxiv 068809; doi: https://doi.org/10.1101/068809
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Highly parallel direct RNA sequencing on an array of nanopores
Daniel R. Garalde, Elizabeth A. Snell, Daniel Jachimowicz, Andrew J. Heron, Mark Bruce, Joseph Lloyd, Anthony Warland, Nadia Pantic, Tigist Admassu, Jonah Ciccone, Sabrina Serra, Jemma Keenan, Samuel Martin, Luke McNeill, Jayne Wallace, Lakmal Jayasinghe, Chris Wright, Javier Blasco, Botond Sipos, Stephen Young, Sissel Juul, James Clarke, Daniel J Turner
bioRxiv 068809; doi: https://doi.org/10.1101/068809

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