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These portions underwent thermal denaturation for 5 min at precisely defined temperatures (37.0, 41.7, 45.8, 49.7, 53.5, 57.4, 61.3, 64.3, 67.0 C) using a MasterCycler EpGradient S thermal cycler (Eppendorf SE, DE)

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It identified negative cases correctly about 75% of the time