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The thermocycler conditions were 95 C for 30 s for 1 cycle, 95 C for 5 s, and 60 C for 10 s for 40 cycles

The association that developed between the response and the independent elements was modeled using the following second-order polynomial equation: In the statistical model, the dependent variable is represented by Y, the independent variables by X i and X j , and the coefficients for the intercept, linear terms, quadratic terms, and interaction terms are denoted by 0 , i , ii , and ij , respectively

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