SARS-CoV-2 passage 5 cat ACE2 rep1
BHK-21 cells were transfected with 0.5 ug of either pUC19 or pGL113 (human), pGL114 (dog), pGL116 (cat), pGL271 (mink), or pGL369 (white-tailed deer) expression plasmids for ACE2. Cells were infected in quadruplicate at a MOI of 0.01 24h later with SARS-CoV-2 Wuhan-Hu-1 (GenBank accession NC_045512.2) bearing the Spike D614G mutation generated from an infectious clone. Supernatants were harvested from each replicate at peak titer and quantified by plaque assay on Vero E6-TMPRSS2-T2A-hACE2 cells. Supernatants were passaged independently for a total of ten passages. RNA was extracted on the unpassaged virus and from passage 5 and 10 viruses using the Zymo Research Quick-RNA Viral kit (R1035). Library preparation was performed using an Illumina COVIDseq assay kit with ARTIC V3 primers. Paired-end 2x150bp sequencing was performed on the Illumina HiSeq platform.