Table 2 Cycling conditions for amplification of the PCR primary and secondary round
PCR conditions | |
|---|---|
Primary round PCR programme: 3Â h | Nested round PCR programme: 3Â h |
1. Initial denaturation—94 °C × 5 m | 1. Initial denaturation—94 °C × 5 m |
2. 40 Cycles—94 °C × 30 s, 52.5 °C × 90 s, 68 °C × 90 s | 2. 40 Cycles—94 °C × 30 s, 60 °C × 90 s, 72 °C × 90 s |
3. Final elongation—68 °C × 10 m | 3. Final elongation—72 °C × 10 m |
4. Hold at 4 °C | 4. Hold at 4 °C |