Table 4 Comparison of the single-primer and double-primer PCR reactions.
| Single-primer PCR | Double-primer PCR | |
|---|---|---|
| Number of reactions | 2 (1 for each primer) | 1 |
| Plasmid template | ~500 ng | 5–50 ng |
| Primers |
Forward OR reverse (each in a separate tube) | Forward AND reverse together |
| Primer-primer annealing during PCR | - | + |
| Number of cycles | 30 | 12–18 |
| Fold amplification of DNA | 30 | 4,096–262,144 |
| Probability of mutation during PCR | Low* | High* |
| DNA polymerase | Pwo | PfuTurbo |