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Nickase-dependent isothermal DNA amplification

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DOI: 10.4236/abb.2013.44070    4,469 Downloads   7,464 Views   Citations
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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 105 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.

Conflicts of Interest

The authors declare no conflicts of interest.

Cite this paper

He, Y. and Jiang, T. (2013) Nickase-dependent isothermal DNA amplification. Advances in Bioscience and Biotechnology, 4, 539-542. doi: 10.4236/abb.2013.44070.

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