Abstract
With the use of polymerases having 3′ to 5′ exonuclease activity and 3′ phosphorothioate-modified allelespecific primers, we recently devised a SNP-operated on/off switch controlling DNA polymerization. One advantage of this novel on/off switch is its adaptability to arrayed primer extension. To further expand its application in genetic analysis, the new on/off switch was evaluated in discrimination of the match/mismatch status of single nucleotides upstream from the primer 3′ terminal. A set of seven amplicons was developed with the templates differing from each other by a single nucleotide. Using this set of amplicons, the new on/off switch was shown to be able to efficiently discriminate single nucleotide polymorphisms from the primer 3′ terminus to the −6 position from the primer 3′ terminus. These data, illustrating the broad single nucleotide discrimination ability of this novel on/off switch, explain why the SNP-operated on/off switch is powerful in SNP analysis, and also indicate useful applications to genetic analysis additional to SNP assay. First, these data broaden the application of the novel on/off switch in the analysis of mutations other than SNPs. Second, it raises a nucleotide-walking algorithm suitable for de novo array-based sequencing analysis.
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Li, K., Zhang, J., Chen, L. et al. Superb nucleotide discrimination by a novel on/off switch for DNA polymerization and its applications. Mol Biotechnol 29, 93–100 (2005). https://doi.org/10.1385/MB:29:2:093
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DOI: https://doi.org/10.1385/MB:29:2:093