| 摘要 |
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 [ DELTA G] effect); or the ligase may itself discriminate between perfect matches and mismatches ( DELTA G or kinetic effect); or some combination of these and other factors.
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