Table 2.
Uracil content measured from DNA samples synthesized by Taq polymerase in the presence of different dUTP/dNTP ratio
| dUTP/dNTP ratio (%) | Deoxyuridine measured (1/million dNMP) |
||
|---|---|---|---|
| Based on 3H -labeled deoxyuridine detection | Based on real-time PCR reactions |
||
| 544 bp amplicon | 1057 bp amplicon | ||
| 0 | 0 | 0 | 0 |
| 0.1 | 233 ± 35 | n.d. | 361 ± 2 |
| 0.2 | 488 ± 32 | 146 ± 463 | 933 ± 163 |
| 0.4 | 1125 ± 68 | n.d. | 1465 ± 43 |
| 0.6 | n.d. | 2587 ± 391 | 2070 ± 76 |
| 0.8 | 2205 ± 148 | n.d. | 2570 ± 4 |
| 1 | 2867 ± 693 | 3538 ± 194 | 3806 ± 13 |
| 2 | 6000 ± 563 | 6301 ± 528 | 5166 ± 8 |
| 3 | 8864 ± 426 | 10 861 ± 346 | n.d. |
| 4 | 12 591 ± 487 | 13 107 ± 209 | n.d. |
| 5 | 15 966 ± 1065 | 13 878 ± 494 | n.d. |
For radioisotopic measurements, 3H labeled dUTP was added to the dNTP pool. First panel: values measured by tritium activity, second panel: values measured by Pfu DNA polymerases amplifying 544 bp in the real-time PCR reaction, third panel: values measured by Pfu DNA polymerases amplifying 1057 bp in the real-time PCR reaction. Values represent mean ± standard error of mean, from 3 to 10 independent measurements, n.d., not determined.