Table 3 Optimization of primer and probe concentration and annealing temperature.
From: Establishment and validation of a simple and accurate qPCR detection method for Haemophilus parasuis
Factor | Parameters | Plasmid concentration(copies/µL) | ||
|---|---|---|---|---|
3.16 × 105 | 3.16 × 104 | 3.16 × 103 | ||
Primer concentration (nM) | 50 | 22.79 ± 0.16 | 27.58 ± 0.54 | 30.48 ± 0.01 |
100 | 22.67 ± 0.25 | 27.25 ± 0.08 | 30.38 ± 0.07 | |
150 | 22.43 ± 0.03 | 26.94 ± 0.06 | 29.96 ± 0.03 | |
Primer concentration(nM) | 50 | 23.41 ± 0.09 | 26.47 ± 0.20 | 30.34 ± 0.08 |
100 | 22.61 ± 0.16 | 25.91 ± 0.05 | 29.66 ± 0.05 | |
150 | 22.67 ± 0.02 | 25.90 ± 0.10 | 29.49 ± 0.05 | |
Annealing temperature (℃) | 58 | 24.20 ± 0.03 | 27.70 ± 0.11 | 31.03 ± 0.11 |
60 | 24.08 ± 0.01 | 27.64 ± 0.09 | 30.91 ± 0.11 | |
62 | 24.17 ± 0.01 | 27.64 ± 0.17 | 31.08 ± 0.01 | |
