| 摘要 |
<p>The invention relates to methods using a modified DNA ligase which increases the discrimination of perfect matches from mismatches for complementary polynucleotides. The modified ligase enhances discrimination in a number of ways, for example, the ligase may increase the difference in the on rates and/or the off rates between a perfect match product and a mismatch product (a kinetic effect); or the ligase may increase the binding energy difference between a perfect match and a mismatch (a free energy [ΔG] effect); or the ligase may itself discriminate between perfect matches and mismatches (ΔG or kinetic effect); or some combination of these and other factors.</p> |
