Table 2.
Error rate values for six PCR enzymes included in this study are presented. Each enzyme was used in two independent PCR reactions. Average doublings/PCR reaction (d) is the average of doubling values for each of the 94 PCR reactions in one plate, where doublings are calculated from the formula 2d = (ng DNA after PCR/ng DNA input). Error rate (f) is calculated as f = n/S (target size × d), where n is the number of mutations observed for all clones that were sequenced and the (target size × d) for each target that was cloned.
| Enzyme | Expt. | Avg. doublings/PCR reaction | Number of clones sequenced | Total bp sequenced | Number of mutations observed | Error rate |
|---|---|---|---|---|---|---|
| Taq | 1 | 20.5 ± 1.2 | 65 | 8.8 × 104 | 54 | 3.0 × 10−5 |
| 2 | 16.7 ± 0.7 | 37 | 4.7 × 104 | 45 | 5.6 × 10−5 | |
|
| ||||||
| AccuPrime-Taq | 1 | 17.0 ± 1.2 | 75 | 1.0 × 105 | 18 | 1.0 × 10−5 |
| 2 | 16.9 ± 0.6 | N.D. | N.D. | N.D. | N.D. | |
|
| ||||||
| KOD | 1 | 20.8 ± 1.5 | 70 | 1.0 × 105 | 16 | 7.6 × 10−6 |
| 2 | 17.6 ± 0.8 | N.D. | N.D. | N.D. | N.D. | |
|
| ||||||
| Pfu (cloned) | 1 | 16.5 ± 1.1 | 151 | 2.0 × 105 | 9 | 2.8 × 10−6 |
| 2 | 12.0 ± 1.8 | N.D. | N.D. | N.D. | N.D. | |
|
| ||||||
| Phusion | 1 | 21.0 ± 1.9 | 175 | 2.4 × 105 | 13 | 2.6 × 10−6 |
| 2 | 16.6 ± 1.1 | N.D. | N.D. | N.D. | N.D. | |
|
| ||||||
| Pwo | 1 | 22.5 ± 1.2 | 170 | 2.4 × 105 | 13 | 2.4 × 10−6 |
| 2 | 17.6 ± 0.6 | N.D. | N.D. | N.D. | N.D. | |
N.D.: not determined.