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

Tyson, J. R.; James, P.; Stoddart, D.; Sparks, N.; Wickenhagen, A.; Hall, G.; Choi, J. H.; Lapointe, H.; Kamelian, K.; Smith, A. D.; Prystajecky, N.; Goodfellow, I.; Wilson, S. J.; Harrigan, R.; Snutch, T. P.; Loman, N. J.; Quick, J.

2020-09-04 genomics
10.1101/2020.09.04.283077 bioRxiv
Show abstract

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 {pound}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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