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Table 1 Optimized RT-PCR program using Quant Studio 3

From: One-step real-time multiplex reverse transcription-polymerase chain reaction assay with melt curve analysis for detection of potato leafroll virus, potato virus S, potato virus X, and potato virus Y

Process RTc PCR (40cycle) Melting curve analysis
Subprocess cDNA synthesis Denature Denature Annealing-Elongation Denature Annealing Meltingd Hold
Tmpa 50 °C 95 °C 95 °C 58 °C 95 °C 60 °C 0.03 °C/s 95 °C
Time 5 min 10 s 5 s 31 s 15 s 15 s   1 s
  1. aTemperature
  2. bMeasure fluorescent intensity in the steps marked with black dot ()
  3. cReverse transcription
  4. dTemperature was increased 0.03 °C per second in this step to dissociate dsDNA gradually