Recent efforts toward the development of electrochemical biosensors have emphasized the incorporation of nanomaterials, such as carbon nanotubes, metal nanoparticles, and graphene into the electrode. This incorporation, particularly of graphene, significantly improves the sensor sensitivity and selectivity.1,2 Graphene, a single-atom-thick twodimensional sp 2 carbon network material, serves as a good support for biomolecules due to its flat surface, large surface area, and rich π-conjugation structure.3-7 The combination of graphene and DNA has attracted much interest in the biosensor field. Recently, the interaction between hairpin DNA (hp-DNA) and graphene surface have been used to detect single nucleotide polymorphisms (SNP).5,8 Hp-DNA is a secondary DNA structure in which two regions of the same DNA strand form base-pairs in a double helix capped by an unpaired loop. The presence of the unpaired loop permits hp-DNA to readily adsorb onto graphene surfaces.The development of efficient methods for detection of trace Hg 2+ is of significant importance for human health and the environment because Hg 2+ is an extremely toxic heavy metal ion. Numerous strategies including optical and electrochemical methods for the detection of Hg 2+ have been developed.9 Among them, electrochemical impedance spectroscopy (EIS) as a label-free method, is highly attractive for biosensing assay due to its high sensitivity and simplicity.
10Herein, we report an Hg 2+ detection on the reduced graphene oxide (RGO) surfaces modified with hp-DNA using EIS. The RGO-modified electrode surface was prepared by the chemical reduction of layer-by-layer (LbL) assembled graphene oxide (GO) and polyethyleneimine (PEI). The hp-DNA used here as the probe DNA included a loop region containing several thymine bases. The hp-DNA was immobilized onto the RGO-modified indium-tin-oxide (ITO) electrode surface. Hg 2+ may be captured between thyminethymine (T-T) mismatched base pairs to form a duplex structure stabilized by covalent bonds between Hg 2+ and the thymine residues (T-Hg11 which results in removal of hp-DNA from graphene surface, as shown in Figure 1(a).EIS was used to monitor the interfacial changes in the electrode surface after exposure to an Hg 2+ solution. The charge transfer resistance of a redox couple was recorded using the RGO-modified ITO electrode surface after exposure to the probe hp-DNA and the subsequent interactions with Hg 2+ in the buffer solution. To the best of our knowledge, this is the first report of using RGO-modified surface for the electrochemical impedimetric detection of Hg
2+. Furthermore, Hg 2+ detection using hairpin DNA immobilized on the RGOmodified electrode surface has never been reported.GO was observed by atomic force microscopy ( Figure 1(b)), which confirms that individual single-layer GO sheets with a thickness of ~1 nm. GO is readily dispersed in water due to its intrinsic oxygen-containing groups, including phenol, carbonyl, and epoxide groups.12 The presence of these groups enables the easy adsorption ...