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A Novel Low Temperature PCR Assured High-Fidelity DNA Amplification

文献类型：期刊论文

作者Hong, GF; Lee, SH; Ge, SC; Zhou, SX
刊名INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
出版日期2013
卷号14期号:6页码:12853-12862
关键词low temperature PCR high-fidelity DNA amplification LoTemp PCR high processivity
通讯作者Hong, GF (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, State Key Lab Mol Biol, 320 Yueyang Rd, Shanghai 200031, Peoples R China.,gfhong@sibcb.ac.cn ; shlee01@snet.net ; scge@sibcb.ac.cn ; sxzhou@sibcb.ac.cn
英文摘要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 degrees C instead of the conventional 94-96 degrees 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.
学科主题Biochemistry & Molecular Biology; Chemistry
类目[WOS]Biochemistry & Molecular Biology ; Chemistry, Multidisciplinary
关键词[WOS]POLYMERASE-CHAIN-REACTION ; PROCESSIVITY ; MUTATIONS ; ARTIFACTS ; TESTS
收录类别SCI
语种英语
WOS记录号WOS:000320772500121
版本出版稿
源URL[http://202.127.25.143/handle/331003/406]  
专题上海生化细胞研究所_上海生科院生化细胞研究所
推荐引用方式
GB/T 7714		 Hong, GF,Lee, SH,Ge, SC,et al. A Novel Low Temperature PCR Assured High-Fidelity DNA Amplification[J]. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES,2013,14(6):12853-12862.
APA Hong, GF,Lee, SH,Ge, SC,&Zhou, SX.(2013).A Novel Low Temperature PCR Assured High-Fidelity DNA Amplification.INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES,14(6),12853-12862.
MLA Hong, GF,et al."A Novel Low Temperature PCR Assured High-Fidelity DNA Amplification".INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES 14.6(2013):12853-12862.

入库方式： OAI收割

来源：上海生物化学与细胞生物学研究所

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