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A 25 L reaction mixture was prepared, comprising 12.5 L PCR Master Mix (2X) (Thermo Scientific), 0.8 L of 10 mM forward primer, 0.8 L of 10 mM reverse primer, 2 L MT-DNA (20 ng/L), and 8.9 L nuclease-free water

Context for readings: 35 dB: Whisper quiet (bedroom acceptable for light sleepers) 45 dB: Quiet conversation (living room acceptable) 55 dB: Normal conversation volume (daytime acceptable) 65+ dB: Loud, disruptive (not acceptable for occupied spaces) Test 4: Power Consumption Measurement I measure actual wattage draw on every fan speed over a 24-hour period, then calculate annual energy costs at $0.15/kWh (U.S