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

A Novel Low Temperature PCR Assured High-Fidelity DNA Amplification

1,†,* , 2,†
,
1
 and
1
1 State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yueyang Road, Shanghai 200031, China 2 Milford Hospital and Milford Molecular Laboratory, 2044 Bridgeport Avenue, Milford, CT 06460, USA These authors contributed equally to this work.
* Author to whom correspondence should be addressed.
Received: 17 May 2013 / Revised: 5 June 2013 / Accepted: 7 June 2013 / Published: 20 June 2013
(This article belongs to the Section Molecular Diagnostics)
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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 low temperature PCR; high-fidelity DNA amplification; LoTemp PCR; high processivity
This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

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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.

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