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

By John R. Tyson, P. James, D. Stoddart, N. Sparks, Arthur Wickenhagen, G. Hall, Ji‐Hyun Choi, H. Lapointe, K. Kamelian, Andrew D. Smith and 7 more. bioRxiv, 2020

357 citations recorded by the source corpus.

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.

Subfields: Infectious Diseases, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/8b2b0cb519fa1dcfe872c0d679ebe48ec06cea6c