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. 2023 Jun 9;4(2):102347. doi: 10.1016/j.xpro.2023.102347

PCR cycling conditions

Steps Temperature Time Cycles
Initial Denaturation 98°C 30 s 1
Denaturation 98°C 10 s 35 cycles
Annealing 67°C 30 s
Extension 72°C 30 s
Final extension 72°C 2 min 1
Hold 4°C forever