Title
Free-Labelled Fluorescent Method for ATP Detection Assisted by T4 DNA Ligase
Author
Hongyun Huang, Shui Fu, Yan Huang, JN Tian, Yanchun Zhao, Shulin Zhao
Year
2017
Journal
Analytical Methods
Abstract
A facile, highly selective biosensor was used for ATP quantitative analysis. Here we introduced two hairpins as matrix. While Exo III digested the duplexes of them, the blocked G-quadruplex released. Then N-Methyl Mesoporphyrin IX (NMM) integrated with G-quadruplex to produce a strong fluorescence. But when ATP was added, it triggered the T4 DNA ligase to link the two hairpins. As a result, Exo III couldn’t hydrolyze the hairpins leading to a low signal out. With this method, limit of detection reached to 3.75 nM, linear range was from 5 nM to 300 nM for ATP. This method has good selectivity and specificity without tedious treatment and label process.
Instrument
J-810
Keywords
Circular dichroism, DNA structure, Biochemistry