Table 1 Nested PCR protocol
Step | Nested 1 PCR | Nested 2 PCR | ||
|---|---|---|---|---|
Primers: PLU1 & PLU5 | Primers: PLU3 & PLU Cal 2 | |||
Product size: ~ 1640 bp | Product size: ~ 1500 bp | |||
Temperature (°C) | Time | Temperature (°C) | Time | |
Initialization | 95 | 5Â min | 95 | 5Â min |
Replication (40 cycles) | ||||
 Denaturation | 96 | 5 s | 96 | 5 s |
 Annealing | 55 | 5 s | 55 | 5 s |
 Extension | 68 | 50 s | 68 | 45 s |
Final extension | 72 | 1Â min | 72 | 1Â min |
Final hold | 4 | ∞ | 4 | ∞ |