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m assay. This is unavoidable and cannot be circumvented in temperature scan-based assays. Heating was applied at a rate of 1 °C/min in the 50-99 °C temperature range, with optical and temperature data sampling at 100 ms intervals. A separate experiment was carried out using buffer alone, and this baseline was subtracted from each DNA melting curve before data treatment. Experiments were conducted in triplicate.
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