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A Novel Low Temperature PCR Assured High-Fidelity DNA Amplification
AbstractAs previously reported, a novel low temperature (LoTemp) polymerase chain reaction (PCR) catalyzed by a moderately heat-resistant (MHR) DNA polymerase with a chemical-assisted denaturation temperature set at 85 °C instead of the conventional 94–96 °C can achieve high-fidelity DNA amplification of a target DNA, even after up to 120 PCR thermal cycles. Furthermore, such accurate amplification is not achievable with conventional PCR. Now, using a well-recognized L1 gene segment of the human papillomavirus (HPV) type 52 (HPV-52) as the template for experiments, we demonstrate that the LoTemp high-fidelity DNA amplification is attributed to an unusually high processivity and stability of the MHR DNA polymerase whose high fidelity in template-directed DNA synthesis is independent of non-existent 3'–5' exonuclease activity. Further studies and understanding of the characteristics of the LoTemp PCR technology may facilitate implementation of DNA sequencing-based diagnostics at the point of care in community hospital laboratories.
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Hong, G.; Lee, S.H.; Ge, S.; Zhou, S. A Novel Low Temperature PCR Assured High-Fidelity DNA Amplification. Int. J. Mol. Sci. 2013, 14, 12853-12862.View more citation formats
Hong G, Lee SH, Ge S, Zhou S. A Novel Low Temperature PCR Assured High-Fidelity DNA Amplification. International Journal of Molecular Sciences. 2013; 14(6):12853-12862.Chicago/Turabian Style
Hong, Guofan; Lee, Sin H.; Ge, Shichao; Zhou, Shaoxia. 2013. "A Novel Low Temperature PCR Assured High-Fidelity DNA Amplification." Int. J. Mol. Sci. 14, no. 6: 12853-12862.