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

Autor:	Hope R. Lapointe, Phillip James, Nicholas J. Loman, Andrew D. Smith, Richard Harrigan, Joshua Quick, Kimia Kamelian, Natalie Sparks, Ji Hyun Choi, Terrance P. Snutch, Natalie Prystajecky, Ian Goodfellow, Sam J. Wilson, Grant Hall, Arthur Wickenhagen, David Stoddart, John R. Tyson
Rok vydání:	2020
Předmět:	Nanopore Computer science Library preparation Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Multiplex polymerase chain reaction Computational biology Primer (molecular biology) Genome health care economics and organizations Article DNA sequencing
Zdroj:	bioRxiv article-version (status) pre article-version (number) 1
DOI:	10.1101/2020.09.04.283077
Popis:	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.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::bee9bd9c9508381a9847f35685cf3e1c https://doi.org/10.1101/2020.09.04.283077 Zobrazit plný text záznamu