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Improvements to the ARTIC multiplex PCR method for SARS-CoV-2 genome sequencing using nanopore

Genome sequencing has been widely deployed to study the evolution of SARS-CoV-2 with more than 90,000 genome sequences uploaded to the GISAID database. We published a method for SARS-CoV-2 genome sequencing (https://www.protocols.io/view/ncov-2019-sequencing-protocol-bbmuik6w) online on January 22,...

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Autores principales: Tyson, John R, James, Phillip, Stoddart, David, Sparks, Natalie, Wickenhagen, Arthur, Hall, Grant, Choi, Ji Hyun, Lapointe, Hope, Kamelian, Kimia, Smith, Andrew D, Prystajecky, Natalie, Goodfellow, Ian, Wilson, Sam J, Harrigan, Richard, Snutch, Terrance P, Loman, Nicholas J, Quick, Joshua
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7480024/
https://www.ncbi.nlm.nih.gov/pubmed/32908977
http://dx.doi.org/10.1101/2020.09.04.283077
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author Tyson, John R
James, Phillip
Stoddart, David
Sparks, Natalie
Wickenhagen, Arthur
Hall, Grant
Choi, Ji Hyun
Lapointe, Hope
Kamelian, Kimia
Smith, Andrew D
Prystajecky, Natalie
Goodfellow, Ian
Wilson, Sam J
Harrigan, Richard
Snutch, Terrance P
Loman, Nicholas J
Quick, Joshua
author_facet Tyson, John R
James, Phillip
Stoddart, David
Sparks, Natalie
Wickenhagen, Arthur
Hall, Grant
Choi, Ji Hyun
Lapointe, Hope
Kamelian, Kimia
Smith, Andrew D
Prystajecky, Natalie
Goodfellow, Ian
Wilson, Sam J
Harrigan, Richard
Snutch, Terrance P
Loman, Nicholas J
Quick, Joshua
author_sort Tyson, John R
collection PubMed
description Genome sequencing has been widely deployed to study the evolution of SARS-CoV-2 with more than 90,000 genome sequences uploaded to the GISAID database. We published a method for SARS-CoV-2 genome sequencing (https://www.protocols.io/view/ncov-2019-sequencing-protocol-bbmuik6w) online on January 22, 2020. This approach has rapidly become the most popular method for sequencing SARS-CoV-2 due to its simplicity and cost-effectiveness. Here we present improvements to the original protocol: i) an updated primer scheme with 22 additional primers to improve genome coverage, ii) a streamlined library preparation workflow which improves demultiplexing rate for up to 96 samples and reduces hands-on time by several hours and iii) cost savings which bring the reagent cost down to £10 per sample making it practical for individual labs to sequence thousands of SARS-CoV-2 genomes to support national and international genomic epidemiology efforts.
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spelling pubmed-74800242020-09-10 Improvements to the ARTIC multiplex PCR method for SARS-CoV-2 genome sequencing using nanopore Tyson, John R James, Phillip Stoddart, David Sparks, Natalie Wickenhagen, Arthur Hall, Grant Choi, Ji Hyun Lapointe, Hope Kamelian, Kimia Smith, Andrew D Prystajecky, Natalie Goodfellow, Ian Wilson, Sam J Harrigan, Richard Snutch, Terrance P Loman, Nicholas J Quick, Joshua bioRxiv Article Genome sequencing has been widely deployed to study the evolution of SARS-CoV-2 with more than 90,000 genome sequences uploaded to the GISAID database. We published a method for SARS-CoV-2 genome sequencing (https://www.protocols.io/view/ncov-2019-sequencing-protocol-bbmuik6w) online on January 22, 2020. This approach has rapidly become the most popular method for sequencing SARS-CoV-2 due to its simplicity and cost-effectiveness. Here we present improvements to the original protocol: i) an updated primer scheme with 22 additional primers to improve genome coverage, ii) a streamlined library preparation workflow which improves demultiplexing rate for up to 96 samples and reduces hands-on time by several hours and iii) cost savings which bring the reagent cost down to £10 per sample making it practical for individual labs to sequence thousands of SARS-CoV-2 genomes to support national and international genomic epidemiology efforts. Cold Spring Harbor Laboratory 2020-09-04 /pmc/articles/PMC7480024/ /pubmed/32908977 http://dx.doi.org/10.1101/2020.09.04.283077 Text en https://creativecommons.org/licenses/by/4.0/It is made available under a CC-BY 4.0 International license (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Tyson, John R
James, Phillip
Stoddart, David
Sparks, Natalie
Wickenhagen, Arthur
Hall, Grant
Choi, Ji Hyun
Lapointe, Hope
Kamelian, Kimia
Smith, Andrew D
Prystajecky, Natalie
Goodfellow, Ian
Wilson, Sam J
Harrigan, Richard
Snutch, Terrance P
Loman, Nicholas J
Quick, Joshua
Improvements to the ARTIC multiplex PCR method for SARS-CoV-2 genome sequencing using nanopore
title Improvements to the ARTIC multiplex PCR method for SARS-CoV-2 genome sequencing using nanopore
title_full Improvements to the ARTIC multiplex PCR method for SARS-CoV-2 genome sequencing using nanopore
title_fullStr Improvements to the ARTIC multiplex PCR method for SARS-CoV-2 genome sequencing using nanopore
title_full_unstemmed Improvements to the ARTIC multiplex PCR method for SARS-CoV-2 genome sequencing using nanopore
title_short Improvements to the ARTIC multiplex PCR method for SARS-CoV-2 genome sequencing using nanopore
title_sort improvements to the artic multiplex pcr method for sars-cov-2 genome sequencing using nanopore
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7480024/
https://www.ncbi.nlm.nih.gov/pubmed/32908977
http://dx.doi.org/10.1101/2020.09.04.283077
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