| Parameter | Profile for first round One-Step RT-PCR for VP7 and VP4 target genes | Profile for second round semi-nested multiplex PCR for VP7 G-typing | Profile for second round semi-nested multiplex PCR for VP4 P-typing | ||||||
|
| Temp. (˚C) | Time (min) | Cycle | Temp. (˚C) | Time (min) | Cycle | Temp. (˚C) | Time (min) | Cycle |
| RT | 50 | 15' | 01 | NA | NA | NA | NA | NA | NA |
| PCR Activation | 95 | 02' | 01 | 94 | 02' | 01 | 94 | 02' | 01 |
| Denaturation | 94 | 0.30'' | 40 | 94 | 01' | 40 | 94 | 01' | 40 |
| Annealing | 45 | 0.45'' | 40 | 45 | 0.45'' | 40 | 45 | 01' | 40 |
| Extension | 72 | 01' | 40 | 72 | 01' | 40 | 72 | 01' | 40 |
| Final Extension | 72 | 07' | 01 | 72 | 07' | 01 | 72 | 07' | 01 |
| Hold | 04 | ∞ | - | 04 | ∞ | - | 04 | ∞ | - |