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Nickase-dependent isothermal DNA amplification
Department of Biochemistry, Institute of Biotechnology, University of South China, Hengyang, China
Department of Genetics, Institute of Life Science & Technology, Huazhong University of Science & Technology, Wuhan, China
- 1 Department of Biochemistry, Institute of Biotechnology, University of South China, Hengyang, China
- 2 Department of Genetics, Institute of Life Science & Technology, Huazhong University of Science & Technology, Wuhan, China
Advances in Bioscience and Biotechnology·Volume 04 (2013)·Pages 539–542·Published 11 April 2013·DOI10.4236/abb.2013.44070
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Abstract
We developed a nicking endonuclease dependent DNA amplification (NDA), using Nt.BstNBI to catalyze single-stranded nick on double-stranded DNA, and Bst DNA polymerase to make extension while sealing the nick and displacing the downstream strand. The displaced single-stranded DNA thereby serves as template for primers hybridization and extension, resulting in exponential synthesis of target DNA under isothermal condition. Over 10 5 folds target DNA amplification can be achieved in 30 minutes, generating DNA product suitable for both diagnosis and DNA cloning. This NDA strategy does not require thermal cycling or prerequisite nucleotides modification, making it suitable for application in the field and at the point-of-care.
KeywordsDNA AmplificationDNA PolymeraseIsothermalNickaseStrand Displacement
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