Table 8. Citrus tatter leaf virus RT-qPCR assay validated for robustness.
| Isolates | Experiment | CTLV RT-qPCR Cq Value | ||||
|---|---|---|---|---|---|---|
| Optimum1 | Annealing Temperature2 | Pipetting Errors2 | ||||
| 58°C / 12 μL | -2°C | +2°C | -2 μL | +2 μL | ||
| IPPN122 | TH2986-48 | 22.16 ± 0.05 | 29.59 ± 0.72 | 30.18 ± 0.38 | 30.41 ± 0.25 | 31.62 ± 2.04 |
| TL100 | TL100A | 24.14 ± 0.06 | 23.62 ± 0.06 | 24.29 ± 0.38 | 24.57 ± 0.08 | 23.93 ± 0.35 |
| TL101 | TL101A | 21.85 ± 0.10 | 21.14 ± 0.02 | 21.49 ± 0.09 | 21.45 ± 0.13 | 21.12 ± 0.27 |
| TL103 | 3288–1 | 24.71 ± 0.10 | 25.82 ± 0.25 | 25.81 ± 0.07 | 26.32 ± 0.06 | 26.01 ± 0.09 |
| TL112 | 3291–9 | 22.75 ± 0.10 | 23.44 ± 0.13 | 23.46 ± 0.02 | 23.82 ± 0.35 | 24.61 ± 2.18 |
| TL113 | 3291–10 | 22.42 ± 0.06 | 22.28 ± 0.10 | 22.04 ± 0.11 | 22.29 ± 0.11 | 21.47 ± 0.15 |
| TL115 | 3170–1 | 23.22 ± 0.10 | 24.66 ± 0.15 | 25.47 ± 0.25 | 25.19 ± 0.05 | 25.29 ± 0.10 |
Abbreviations: Cq: quantitative cycle.
1Optimum setup was using the conditions validated and optimized in this study including volume, primer probe concentrations, annealing temperature, etc. And the reactions were run on ThermoFisher Scientific QuantStudio 12K Flex Real-Time PCR System.
2The RT-qPCR reactions were setup with same concentration of primers and probe and using Bio-Rad iTaq™ Universal Probes One-Step Kit per manufacturer's instruction. The reactions were run on Bio-Rad CFX-96 Real-Time PCR Detection System.