TITLE:
Nickase-dependent isothermal DNA amplification
AUTHORS:
Yan He, Tao Jiang
KEYWORDS:
DNA Amplification; DNA Polymerase; Isothermal; Nickase; Strand Displacement
JOURNAL NAME:
Advances in Bioscience and Biotechnology,
Vol.4 No.4,
April
19,
2013
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.