SARS-CoV-2 sequencing from air sample, Wisconsin
Samples were kept frozen at -70C until total nucleic extraction was performed using 200 L elute and Applied Biosystems MagMAX Viral/Pathogen Nucleic Acid Isolation Kit using ThermoFisher Scientific KingFisher Flex instrument. Real-time RT-PCR setup was performed using 10 L of extract and approved Applied Biosystems TaqPathTM COVID-19 Combo Kit containing three primer/probe sets specific to different SARS-CoV-2 genomic regions (open reading frame 1ab (ORF1ab), spike (S) protein, and nucleocapsid (N) protein-encoding genes) and primers/probes for bacteriophage MS2 which served as internal process control for nucleic acid extraction. RT-PCR assay was performed on Applied Biosystems 7500 Fast Dx Real-Time PCR System according to the TaqPathTM COVID-19 Combo Kit protocol. PCR results were interpreted using the Applied Biosystems COVID-19 Interpretive Software.
C_S72_L001