COVID-19 ARTIC v3 Illumina library construction and sequencing protocol v2
DNA Pipelines R D, Benjamin Farr, Diana Rajan, Emma Dawson, Lesley Shirley, Michael A. Quail, Naomi Park, Nicholas Redshaw, Iraad F. Bronner, Louise Aigrain, Scott Goodwin, Scott Thurston, Stefanie Lensing, Charlotte Beaver, Rachel Nelson, Alex Alderton, Ian Johnston
Abstract
This SOP describes the procedure for generating cDNA from SARS-CoV-2 viral nucleic acid extracts and subsequently producing 400nt amplicons tiling the viral genome using V3 nCov-2019 primers (ARTIC). This is followed by library construction, equivolume pooling of samples and quantitation, prior to sequencing on the Illumina NovaSeq. It is adapted from the nCov-2019 sequencing protocol which can be found here: https://www.protocols.io/view/ncov-2019-sequencing-protocol-bbmuik6w.
Topics & Concepts
Illumina dye sequencingPoolingDNA sequencingComputational biologyAmpliconDeep sequencingCoronavirus disease 2019 (COVID-19)Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)BiologyGenomeVirologyGeneticsComputer sciencePolymerase chain reactionDNAGeneMedicineInfectious disease (medical specialty)DiseasePathologyArtificial intelligenceCOVID-19 diagnosis using AI