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2.4.1. TaqMan fast virus 1-Step master mix (Applied Biosystems, cat. 4,444,434)
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A 20 μL PCR final reaction volume contained 5 μL of 4X Reaction Mix, 0.5 μL of LightMix® Modular SARS and Wuhan CoV E-gene, 9.5 μL of nuclease-free water and 5 μL of sample RNA. The thermocycling conditions consisted of a reverse transcription at 50 °C for 5 min, followed by denaturation at 95 °C for 20 s and 45 cycles of 94 °C for 15 s and 58 °C for 30 s. Fluorescence was read in the FAM channel.
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