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The thermal cycling program consisted of an initial denaturation step at 95 C for 8 min, followed by 40 cycles of 95 C for 15 s, 58.9 C for 30 s, and 72 C for 30 s, concluding with a melting curve analysis

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7 A big factor in the prevalence of this high mortality rate is that alcohol impairs peoples decision-making and judgment abilities

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