Int. J. Mol. Sci. 2013, 14(6), 12853-12862; doi:10.3390/ijms140612853

A Novel Low Temperature PCR Assured High-Fidelity DNA Amplification

1,†,* email, 2,†email, 1email and 1email
Received: 17 May 2013; in revised form: 5 June 2013 / Accepted: 7 June 2013 / Published: 20 June 2013
(This article belongs to the Section Molecular Diagnostics)
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract: As 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.
Keywords: low temperature PCR; high-fidelity DNA amplification; LoTemp PCR; high processivity
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MDPI and ACS Style

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.

AMA Style

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.

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