A simple and isothermal ligase-based amplification approach based on a ligation-activated cleavage reaction†
Abstract
We develop a simple and isothermal ligase-based cyclic amplification approach on the basis of a ligation-triggered duplex-specific nuclease (DSN)-mediated cleavage reaction without the involvement of any thermal cycling and separation procedures. As a proof of concept, we demonstrate the application of this method for the sensitive detection of polynucleotide kinase. Moreover, this method can be extended to the accurate quantification of DNA, proteins and ATP.