Table 4 Protocols used for conventional PCR and RT-PCR
From: Evaluation of real-time PCR targeting the lipL32 gene for diagnosis of Leptospira infection
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 | |||||||||