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In the second step, 7 l cDNA from step-1 was amplified using N gene forward primer (5-CACATTGGCACCCGCAATC-3), N gene reverse primer (5-GAGGAACGAGAAGAGGCTTG-3) and probe (5-FAM-ACTTCCTCAAGGAACAACATTGCCA-QSY-3) using Taqman mastermix (Thermo Fisher)

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