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Protocols used for conventional PCR and RT-PCR
| Stage | PCR | RT-PCR | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| LeptoA/LeptoB | L3/L4 | L4/LPato2 | ||||||||||
| Temp. (°C) | Time (min) | Cycles (n) | Temp. (°C) | Time (min) | Cycles (n) | Temp. (°C) | Time (min) | Cycles (n) | Temp. (°C) | Time (s) | Cycles (n) | |
| Initial denaturation | 93 | 3 | 1 | 93 | 3 | 1 | 93 | 3 | 1 | 600 | ||
| Denaturation | 93 | 1 | 35 | 93 | 1 | 35 | 93 | 1 | 35 | 95 | 15 | 45 |
| Annealing | 60 | 1 | 57 | 1 | 52 | 1 | 53 | 10 | ||||
| Elongation | 72 | 1 | 72 | 1 | 72 | 1 | 72 | 10 | ||||
| Final elongation | 72 | 9 | 1 | 72 | 9 | 1 | 72 | 9 | 1 | 40 | 30 | 1 |
| Hold | 4 | 4 | 4 | |||||||||
