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

By John R Tyson, Phillip James, David Stoddart, Natalie Sparks, Arthur Wickenhagen, Grant Hall, Ji Hyun Choi, Hope Lapointe, Kimia Kamelian, Andrew D Smith, Natalie Prystajecky, Ian Goodfellow, Sam J Wilson, Richard Harrigan, Terrance P Snutch, Nick J Loman, Joshua Quick

Posted 04 Sep 2020
bioRxiv DOI: 10.1101/2020.09.04.283077

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 Statement J.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.

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