ConcatSeq: A method for increasing throughput of single molecule sequencing by concatenating short DNA fragments

Abstract Single molecule sequencing (SMS) platforms enable base sequences to be read directly from individual strands of DNA in real-time. Though capable of long read lengths, SMS platforms currently suffer from low throughput compared to competing short-read sequencing technologies. Here, we presen...

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Autores principales:	Ulrich Schlecht, Janine Mok, Carolina Dallett, Jan Berka
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/4873498f3d0c4d4faaefddfe4d1a87aa
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spelling	oai:doaj.org-article:4873498f3d0c4d4faaefddfe4d1a87aa2021-12-02T12:32:59ZConcatSeq: A method for increasing throughput of single molecule sequencing by concatenating short DNA fragments10.1038/s41598-017-05503-w2045-2322https://doaj.org/article/4873498f3d0c4d4faaefddfe4d1a87aa2017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-05503-whttps://doaj.org/toc/2045-2322Abstract Single molecule sequencing (SMS) platforms enable base sequences to be read directly from individual strands of DNA in real-time. Though capable of long read lengths, SMS platforms currently suffer from low throughput compared to competing short-read sequencing technologies. Here, we present a novel strategy for sequencing library preparation, dubbed ConcatSeq, which increases the throughput of SMS platforms by generating long concatenated templates from pools of short DNA molecules. We demonstrate adaptation of this technique to two target enrichment workflows, commonly used for oncology applications, and feasibility using PacBio single molecule real-time (SMRT) technology. Our approach is capable of increasing the sequencing throughput of the PacBio RSII platform by more than five-fold, while maintaining the ability to correctly call allele frequencies of known single nucleotide variants. ConcatSeq provides a versatile new sample preparation tool for long-read sequencing technologies.Ulrich SchlechtJanine MokCarolina DallettJan BerkaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-10 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Ulrich Schlecht Janine Mok Carolina Dallett Jan Berka ConcatSeq: A method for increasing throughput of single molecule sequencing by concatenating short DNA fragments
description	Abstract Single molecule sequencing (SMS) platforms enable base sequences to be read directly from individual strands of DNA in real-time. Though capable of long read lengths, SMS platforms currently suffer from low throughput compared to competing short-read sequencing technologies. Here, we present a novel strategy for sequencing library preparation, dubbed ConcatSeq, which increases the throughput of SMS platforms by generating long concatenated templates from pools of short DNA molecules. We demonstrate adaptation of this technique to two target enrichment workflows, commonly used for oncology applications, and feasibility using PacBio single molecule real-time (SMRT) technology. Our approach is capable of increasing the sequencing throughput of the PacBio RSII platform by more than five-fold, while maintaining the ability to correctly call allele frequencies of known single nucleotide variants. ConcatSeq provides a versatile new sample preparation tool for long-read sequencing technologies.
format	article
author	Ulrich Schlecht Janine Mok Carolina Dallett Jan Berka
author_facet	Ulrich Schlecht Janine Mok Carolina Dallett Jan Berka
author_sort	Ulrich Schlecht
title	ConcatSeq: A method for increasing throughput of single molecule sequencing by concatenating short DNA fragments
title_short	ConcatSeq: A method for increasing throughput of single molecule sequencing by concatenating short DNA fragments
title_full	ConcatSeq: A method for increasing throughput of single molecule sequencing by concatenating short DNA fragments
title_fullStr	ConcatSeq: A method for increasing throughput of single molecule sequencing by concatenating short DNA fragments
title_full_unstemmed	ConcatSeq: A method for increasing throughput of single molecule sequencing by concatenating short DNA fragments
title_sort	concatseq: a method for increasing throughput of single molecule sequencing by concatenating short dna fragments
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/4873498f3d0c4d4faaefddfe4d1a87aa
work_keys_str_mv	AT ulrichschlecht concatseqamethodforincreasingthroughputofsinglemoleculesequencingbyconcatenatingshortdnafragments AT janinemok concatseqamethodforincreasingthroughputofsinglemoleculesequencingbyconcatenatingshortdnafragments AT carolinadallett concatseqamethodforincreasingthroughputofsinglemoleculesequencingbyconcatenatingshortdnafragments AT janberka concatseqamethodforincreasingthroughputofsinglemoleculesequencingbyconcatenatingshortdnafragments
_version_	1718393914383138816

ConcatSeq: A method for increasing throughput of single molecule sequencing by concatenating short DNA fragments

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