| 摘要 |
High-throughput detection for the interesting base or the mutation site in the nucleic acid sample can be achieved by the linear test probe pairs P1 and P2. The test probe pairs P1 and P2 respectively comprise either of the flanking complementary sequences which are adjacent to the interesting base or the mutation site in the nucleic acid sample. When the test probe pairs P1, P2 are annealed and hybridized to the nucleic acid sample, a gap will be generated at the interesting base or the mutation site position between the probe pairs and the sample. Divide the annealed hybrid sample into four equal reaction systems to which add dATP, dTTP, dCTP, dGTP, respectively. The test probe pairs P1 and P2 will be ligated into one single probe when adding the complementary nucleotide system under the DNA polymerase or ligase. After purified and amplified, the generated single probes are hybridized to the corresponding area in a common oligonucleotide microarray. The generated single probe will give a signal in the hybrid area, and therefore detect and analyze the hybrid signal to determine the base type or the mutation genotype at the detection position. The invention can be applied to the re-sequencing the target nucleic acid sequence, the detection and analysis for the mutation, insertion, or deletion sites of a known nucleic acid sequence, and the genotyping of the pathogenic microorganism.
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