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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,...
Autores principales: | , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cold Spring Harbor Laboratory
2020
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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. |
format | Online Article Text |
id | pubmed-7480024 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Cold Spring Harbor Laboratory |
record_format | MEDLINE/PubMed |
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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