Taqman sars cov 2 assay kit version 2

For the RT-PCR, the TaqMan SARS-CoV-2 Assay Kit version 2 (Thermo Fisher; catalog number CCU002NR) was used, as described previously.⁷ (link) Each reaction contained 6.25 μL TaqPath 1-Step Multiplex Master Mix, NO ROX (Thermo Fisher; catalog number A28523), 1.25 μL TaqMan SARS-CoV-2 Assay Kit version 2 (primers and probes), 1.00 μL TaqMan MS2 Phage, 11.50 μL nuclease-free water, and 5 μL of sample RNA, nontemplate nuclease-free water control, or TaqMan SARS-CoV-2 Control Kitv2 (CCU002NR) positive control. The one-step RT-PCR was executed on a QuantStudio 5 DX real-time PCR-System (Thermo Fisher; catalog number A36324) machine with the following steps: Uracil N-glycosylase incubation (25°C, 2 minutes), reverse transcription (53°C, 10 minutes), activation (95°C, 2 minutes), and 45 cycles of denaturation (95°C, 3 seconds) and annealing/extension (60°C, 30 seconds).