QuantiNova Probe RT-PCR Kit (2500)

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Publication protocol

A non-nested COVID-19-RdRp/Hel real-time RT-PCR assay for SARS-CoV-2 RNA detection was performed using a QuantiNova Probe RT-PCR Kit (QIAGEN, Hilden, Germany), as we previously described [14]. Each 20 μL reaction mixture contained 10 μL of 2× QuantiNova Probe RT-PCR Master Mix, 0.2 μL of QN Probe RT-Mix, 1.6 μL of each 10 μM forward and reverse primer, 0.4 μL of 10 μM probe (Table 3), 1.2 μL of nuclease-free water and 5 μL of TNA. Amplification and detection were performed on a LightCycler 480 II Real-Time PCR System (Roche, Basel, Switzerland). Thermocycling conditions consisted of 10 min at 45 °C for reverse transcription, 5 min at 95 °C for PCR initial activation, and 45 cycles of 5 s at 95 °C and 30 s at 55 °C.

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