This allows adjustments of the magnesium concentration to optimal levels for each reaction. High annealing temperatures tend to minimize nonspecific primer annealing, thereby increasing the amount of specific product and decreasing primer-dimer formation. A total of 25-30 cycles usually is adequate for the visual detection of the sequence of interest. This technique increases PCR specificity and yield of the desired product by keeping the polymerase separate from the rest of the reaction components until a critical temperature is reached. References D'Aquila, R.T. et al .
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