Table 2.
Effect of primer structure and dTTP/Mg2+ on interaction of human PolεCD with template:primer.
| Primer sequence | koff × 10–3 s−1 | kon (mM−1 s−1) | KDa (nM) | |||
|---|---|---|---|---|---|---|
| − | + dTTP | − | + dTTP | − | + dTTP | |
| GCCUGGAGCGC (0) | 21.9 ± 2.2 | – | 336 ± 80 | – | 65.8 ± 8.8 | – |
| GCCUGGAGCG/ddC/ (1)a | 19.1 ± 2.5 | 88.6 ± 11 | 269 ± 38 | 210 ± 15 | 73.0 ± 14 | 424 ± 22 |
| GCCUGGAGCG/ddC/ (3) | 19.8 ± 0.1 | 110 ± 18 | 318 ± 29 | 250 ± 24 | 62.5 ± 6.1 | 439 ± 28 |
| GCCUGGAGCG/ddC/ (5) | 18.2 ± 1.8 | 58.5 ± 1.7 | 264 ± 51 | 269 ± 24 | 69.8 ± 6.4 | 218 ± 13 |
| GCCUGGAGCG/ddC/ (7) | 15.0 ± 2.1 | 22.6 ± 2.7 | 257 ± 31 | 273 ± 22 | 58.6 ± 1.3 | 83 ± 3.3 |
| GCCTGGAGCG/ddC/ (11) | 21.8 ± 3.9 | 31.9 ± 5.6 | 356 ± 7.2 | 264 ± 24 | 61.4 ± 12 | 120 ± 11 |
| GCCTGGAGCG/3dC/ (10)b | 17.1 ± 1 | 54.9 ± 0.2 | 378 ± 27 | 244 ± 13 | 45.6 ± 6.1 | 225 ± 13 |
2′-Deoxy- and 2′,3′-dideoxy- nucleotides are underlined and their quantity is indicated in parenthesis.
The provided values for DNA primers at 0.1 M NaCl are from Table 1.
Data are presented as mean ± SD (n = 3).
a/ddC/—2′,3′-dideoxy-cytosine.
b/3dC/—3′-deoxy-cytosine.