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2.4.2. iTaq™ universal probes one-step kit (Bio-rad, cat. 1,725,141)
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A 20 μL PCR final reaction volume contained 10 μL of 2X iTaq Master Mix, 0.5 μL of LightMix® Modular SARS and Wuhan CoV E-gene, 4.25 μL of nuclease-free water, 0.25 μL iScript Reverse Transcriptase and 5 μL of sample RNA. The thermocycling conditions consisted of a reverse transcription at 50 °C for 20 min, followed by denaturation at 95 °C for 5 min and 45 cycles of 94 °C for 10 s and 58 °C for 30 s. Fluorescence was read in the FAM channel.
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