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marginale was used ( The first reaction mixture comprised 1.2 µl dNTPs (2.5 mM), 0.15 µl Taq polymerase (0.05 U), 1.5 µl reaction buffer IB (1×), 0.6 µl of a solution containing the mixed primers (10 mM), and 10.05 µl sterile ultra-pure water