摘要 |
<p>The present invention involves methods of detecting small mutations in target nucleic acid using the Ligase Chain Reaction (LCRTM). Detectable small mutations include single base deletions, insertions and changes, as well as multiple mutations (deletions, insertions and changes) where the size of the mutation is less than about 15 % of the average probe length. The methods include high concentrations of monovalent cations, especially Na+, K+ and NH¿4?+; and/or relatively high initial mixing temperatures. The methods also include probe designs having deliberate mismatches with respect to target, the mismatches being near the 5' ends. Also claimed are several sets of probe sequences for detection of mutations associated with cystic fibrosis.</p> |