Electrochemical DNA Hybridization Detection Using DNA Cleavage

Abstract

We report the new method for detection of DNA hybridization using enzymatic cleavage. The strategy is based on that S1 nuclease is able to specifically cleave only single strand DNA, but not double strand DNA. The capture probe DNA, thiolated single strand DNA labeled with electroactive ferrocene group, was immobilized on a gold electrode. After hybridization of target DNA of complementary and noncomplementary sequences, nonhybridized single strand DNA was cleaved using S1 nuclease. The difference of enzymatic cleavage on the modified gold electrode was characterized by cyclic voltammetry and differential pulse voltammetry. We successfully applied this method to the sequence-selective discrimination between perfectly matched and mismatched target DNA including a single-base mismatched target DNA. Our method does not require either hybridization indicators or other exogenous signaling molecules which most of the electrochemical hybridization detection systems require.

Label-Free Fluorescent Detection of Ions, Proteins, and Small Molecules Using Structure-Switching Aptamers, SYBR Gold, and Exonuclease I

“…nicking enzymes, 24−26 S1 nuclease,27−29 Exo III, 30−32 Exo I, 33−35 and so on, are widely used in various biosensing techniques to achieve exceptional detection selectivity and sensitivity for DNA detection. Recently, the application of nucleases in biosensing has extended to nonnucleic acid targets by utilizing SSAs as recognition molecules.…”

A novel electrochemical method to detect theophylline utilizing silver ions captured within abasic site-incorporated duplex DNA

“…The result shows that the oxidation peak current quite linearly increase as a function of scan rate, indicating the current completely and exclusively relies on the surface-confined signal produced by the silver ion captured within AP site-incorporated duplex DNA immobilized on the electrode surface (Fig. S2) (Kwon et al, 2008). We also investigated the effect of immersion time of silver nitrate on the current signal.…”

Discovery of the unique self-assembly behavior of terminal suckers-contained dsDNA onto GNP and novel “light-up” colorimetric assay of nucleic acids

“…Accordingly, various DNA sensors based on the electronic technique (Kwon et al, 2008), mass spectrometry (Hofstadler and Griffey, 2001) and optical measurements (Guo et al, 2009;Shalon et al, 1996;Hou et al, 2014) have been raised. However, most of these suffer from the limitations including high-cost instruments, complicated operation or inadequate sensitivity, making them difficult to prevail particularly in the developing world.…”