RT Journal Article
SR Electronic
T1 Improvements to the ARTIC multiplex PCR method for SARS-CoV-2 genome sequencing using nanopore
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.09.04.283077
DO 10.1101/2020.09.04.283077
A1 John R Tyson
A1 Phillip James
A1 David Stoddart
A1 Natalie Sparks
A1 Arthur Wickenhagen
A1 Grant Hall
A1 Ji Hyun Choi
A1 Hope Lapointe
A1 Kimia Kamelian
A1 Andrew D Smith
A1 Natalie Prystajecky
A1 Ian Goodfellow
A1 Sam J Wilson
A1 Richard Harrigan
A1 Terrance P Snutch
A1 Nicholas J Loman
A1 Joshua Quick
YR 2020
UL http://biorxiv.org/content/early/2020/09/04/2020.09.04.283077.abstract
AB 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.Competing Interest StatementJ.R.T., J.Q., and N.J.L. have all received travel expenses and accommodation from Oxford Nanopore Technologies to speak at organised events. P.J. and D.S. are employees of Oxford Nanopore Technologies.