The 25 reaction mixtures contained 0.5 of primer, 2.5 of 10 buffer, 0.5 Taq DNA polymerase (Gibco BRL, Gaithersburg, MD, USA) and 0.5 deoxyribonucleoside triphosphates. The PCR protocol comprised an initial denaturation step at 95 for 15 min, followed by 35 cycles of a denaturation step at 94 for 20 s, a primer annealing step at 50 for 45 s, an extension step at 72 for 30 s and a final step of 72 for 5 min. Amplicons were analyzed by 1.5 agarose gel electrophoresis performed at 4 V in Tris borate EDTA buffer.
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