Each PCR reaction consisted of an initial denaturation step (5 min at 94°C) followed by 39 cycles of PCR (50 s at 94°C, 1 min at annealing temperature depending on the region to be amplified, and 1 min at 72°C) and a final extension step of 5 min at 72°C, or of 10 min if the amplified segment was to be cloned.
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