| 摘要 |
An improved assay for identifying and distinguishing one or more a single nucleotide polymorphisms in one or more target sequences of nucleic acid comprises, in a single-tube reaction system, three or more primers, two of which bind to a target nucleic acid sequence, flanking a SNP, so that the 3'-end of one or more first primers is adjacent to the 5'-end of a second primer, the two primers being selectively ligated and then amplified by a third primer to exponentially produce the complementary strand of the one or more target sequences. The other strand of the one or more target sequences are exponentially amplified by one or more hybridizable probes, each labeled with a different fluorophore, the fluorophore-labeled hybridizable probes being quenched until incorporation into and amplification of target nucleic acid products. Also provided is a method for identifying one or more SNPs in one or more target sequences of nucleic acid in each single through-hole of a nanoliter sampling array, and a kit for such a method containing a nanoliter sampling array chip, primer sequences, and reagents required to selectively ligate primers for amplification of desired target nucleic acid sequences.
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