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Illumina Nextera DNA Flex library construction and sequencing for SARS-CoV-2:Adapting COVID-19 ARTIC protocol v1

Year: 2020
Type: preprint
Abstract: This protocol describes the procedure for generating cDNA from SARS-CoV-2 viral nucleic acid extracts and subsequently producing amplicons tiling the viral genome sequencing. It uses the V3 nCov-2019 primers from the ARTIC network. This is followed by library construction using Nextera Flex, which we found to save 9h of hands on time as compared with original protocol that use TrueSeq for library contstruction. It also describes the pooling of samples and quantitation, prior to sequencing on the Illumina Miseq and NextSeq. It is adapted from the nCov-2019 sequencing protocol from Quick and colleagues, which can be found here:
DOI: 10.17504/protocols.io.bhjgj4jw
Authors: Sureshnee Pillay, Jennifer Giandhari, Houriiyah Tegally, Eduan Wilkinson, Benjamin Chimukangara, Richard Lessells, Yunus Moosa, Inbal Gazy, Maryam Fish, Lavanya Singh, Khulekani Sedwell Khanyile, Vagner Fonseca, Marta Giovanetti, Luiz Carols Alcantara, Tulio de Oliveira
Institutions: Universidade Federal de Minas Gerais, University of Washington, Centre for the AIDS Programme of Research in South Africa, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, University of KwaZulu-Natal
Language: English
Field: Medicine
Open Access status: green
OpenAlex ID: W3040125315
Cites: 1
Cited by: 7
Related to: 10