High Throughput Nanopore Sequencing of SARS-CoV-2 Viral Genomes from Patient Samples

Pater, Adrian A.; Bosmeny, Michael S.; Parasrampuria, Mansi; Eddington, Seth B.; Ovington, Katy N.; White, Adam A.; Barkau, Christopher L.; Metz, Paige E.; Sylvain, Rourke J.; Chilamkurthy, Ramadevi; Yinusa, Abadat O.; Benzinger, Scott W.; Hebert, Madison M.; Gagnon, Keith T.

In late 2019, a novel coronavirus began spreading in Wuhan, China, causing a potentially lethal respiratory viral infection. By early 2020, the novel coronavirus, called SARS-CoV-2, had spread globally, causing the COVID-19 pandemic. The infection and mutation rates of SARS-CoV-2 make it amenable to tracking movement and evolution by viral genome sequencing. Efforts to develop effective public health policies, therapeutics, or vaccines to treat or prevent COVID-19 are also expected to benefit from tracking mutations of the SARS-CoV-2 virus. Here we describe a set of comprehensive working protocols, from viral RNA extraction to analysis using online visualization tools, for high throughput sequencing of SARS-CoV-2 viral genomes using a MinION instrument. This set of protocols should serve as a reliable ‘how-to’ reference for generating quality SARS-CoV-2 genome sequences with ARTIC primer sets and next-generation nanopore sequencing technology. In addition, many of the preparation, quality control, and analysis steps will be generally applicable to other sequencing platforms.