Abstract
DNA ligation is a popular mechanism in analyses of gene sequences. The ligation of pairs of DNA probes upon hybridization to a target nucleic acid sequence creates a contiguous DNA sequence that can be used to infer the presence and nature of target molecules. The ligatable DNA strands can also be coupled to other molecules interacting with targets of interest to increase the range of molecules detectable in ligase-based assays. In ligase-mediated detection of nucleic acid sequences, the requirements for substrate recognition by ligases may be used to distinguish target sequences that differ in even single nucleotide positions. The covalent link that forms between the probe pairs can be used to connect retrievable functions on one probe to detectable groups on the other. Even more usefully, the resulting contiguous DNA strand can be amplified, along with sequence elements that encode the target specificity of the probes. These principles are being applied in a rapidly increasing range of molecular analyses, including high-throughput genotyping, in situ detection of nucleic acid sequences, and highly sensitive protein assays.
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