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  • Title: [Effect of 3' exonuclease activity of polymerase on extension of phosphorothioate-modified primers].
    Author: Guo ZF, Chen LL, Zhang J, Peng CY, Yang XD, Zhang X, He SY, Liao DF, Li K.
    Journal: Zhonghua Yi Xue Yi Chuan Xue Za Zhi; 2003 Aug; 20(4):328-30. PubMed ID: 12903045.
    Abstract:
    OBJECTIVE: To determine whether 3'phosphorothioate-modified-2 terminal mismatched primers can turn off DNA polymerization mediated by Exo(+) polymerase. METHODS: Two-directional primer extension was performed using polymerase with and without 3' exonuclease activity. The effects of unmodified primers and 3' phosphorothioate-modified primers on primer extension were evaluated. RESULTS: Exo(-) polymerase yielded products from matched and mismatched primers regardless of their modification. However, 3' phosphorothioate-modified primers with a single base mismatch at -2 position worked similarly to the terminal (-1) mismatched primers in triggering the novelly reported "off-switch" of Exo(+) polymerase. CONCLUSION: These data suggested that the "off-switch" can be of enormous application in the diagnosis of single gene diseases and in the association studies by single nucleotide polymorphism screening.
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