200 μL of the supernatant was harvested from cells infected with SARS-CoV-2 and mixed with equal volume of lysis buffer provided in the high pure viral RNA kit (Invitrogen) and RNA was extracted according to the kit's instructions. The eluted RNA was subjected to RT-qPCR by using the One Step TB Green PrimeScript RT-qPCR Kit II (Takara) and specific primers targeting the SARS-CoV-2 nucleocapsid protein (NP):
Forward, 5′-TAATCAGACAAGGAACTGATTA-3′;
Reverse, 5′-CGAAGGTGTGACTTCCATG-3′.
RT-qPCR cycling conditions were 42 °C for 5 min, 95 °C for 10 s, and 40 cycles of 95 °C for 5 s, followed by 60 °C for 30 s.
All the SARS-CoV-2 based experiments were performed at the UCLA BSL3 facility.
Forward, 5′-TAATCAGACAAGGAACTGATTA-3′;
Reverse, 5′-CGAAGGTGTGACTTCCATG-3′.
RT-qPCR cycling conditions were 42 °C for 5 min, 95 °C for 10 s, and 40 cycles of 95 °C for 5 s, followed by 60 °C for 30 s.
All the SARS-CoV-2 based experiments were performed at the UCLA BSL3 facility.