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Isothermal DNA amplification in vitro: the helicase-dependent amplification system

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Abstract

Since the development of polymerase chain reaction, amplification of nucleic acids has emerged as an elemental tool for molecular biology, genomics, and biotechnology. Amplification methods often use temperature cycling to exponentially amplify nucleic acids; however, isothermal amplification methods have also been developed, which do not require heating the double-stranded nucleic acid to dissociate the synthesized products from templates. Among the several methods used for isothermal DNA amplification, the helicase-dependent amplification (HDA) is discussed in this review with an emphasis on the reconstituted DNA replication system. Since DNA helicase can unwind the double-stranded DNA without the need for heating, the HDA system provides a very useful tool to amplify DNA in vitro under isothermal conditions with a simplified reaction scheme. This review describes components and detailed aspects of current HDA systems using Escherichia coli UvrD helicase and T7 bacteriophage gp4 helicase with consideration of the processivity and efficiency of DNA amplification.

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Acknowledgments

This work was supported by a grant (20080401034026) from the BioGreen 21 Program, Rural Development Administration, Republic of Korea, a grant from Korea Health 21 R&D Project, Ministry of Health & Welfare, Republic of Korea (A080123), a grant (10032113) from Industrial Technology Development, Ministry of Knowledge Economy, and a grant from the Korea Science & Engineering Foundation (KOSEF) through a general research grant (R01-2008-000-20301-0). K. Park is supported by the second stage of Brain Korea 21. Y.-J. Jeong was supported by the Korea Research Foundation Grant funded by the Korean Government (MOEHRD, Basic Research Promotion Fund) (KRF-2008-313-C00531).

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Authors and Affiliations

  1. Department of Bio and Nanochemistry, Kookmin University, 861-1 Jeongneung-dong, Seongbuk-gu, Seoul, 136-702, Republic of Korea

    Yong-Joo Jeong

  2. Department of Bioscience and Biotechnology, Konkuk University, 1 Hwayang-dong, Gwanjin-gu, Seoul, 143-701, Republic of Korea

    Kkothanahreum Park & Dong-Eun Kim

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  1. Yong-Joo Jeong
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  2. Kkothanahreum Park
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  3. Dong-Eun Kim
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Correspondence to Dong-Eun Kim.

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Jeong, YJ., Park, K. & Kim, DE. Isothermal DNA amplification in vitro: the helicase-dependent amplification system. Cell. Mol. Life Sci. 66, 3325–3336 (2009). https://doi.org/10.1007/s00018-009-0094-3

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  • DOI: https://doi.org/10.1007/s00018-009-0094-3

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