Controlled orientation of DNA in a binary SAM as a key for the successful determination of DNA hybridization by means of electrochemical impedance spectroscopy.

Determination of DNA hybridization at electrode surfaces modified with thiol-tethered single-stranded DNA (ssDNA) capture probes and co-assembled with short-chain thiol derivatives using electrochemical impedance spectroscopy requires a careful design of the electrode/electrolyte interface as well as an in-depth understanding of the processes at the interface during DNA hybridization. The influence of the electrode potential, the ssDNA coverage, the ionic strength, the nature of the thiol derivative and especially the Debye length are shown to have a significant impact on the impedance spectra. A mixed monolayer comprising--in addition to the ssDNA capture probe--both mercaptohexanol (MCH) and mercaptopropionic acid (MPA) is suggested as an interface design which allows a high efficiency of the DNA hybridization concomitantly with a reliable modulation of the charge-transfer resistance of the electrode upon hybridization.

[1]  Nicole Jaffrezic-Renault,et al.  Impedance spectroscopy and affinity measurement of specific antibody–antigen interaction , 2006 .

[2]  Jean Sturm,et al.  Persistence Length of Single-Stranded DNA , 1997 .

[3]  Bobby Pejcic,et al.  Impedance spectroscopy: Over 35 years of electrochemical sensor optimization , 2006 .

[4]  N. Pourmand,et al.  Label-Free Impedance Biosensors: Opportunities and Challenges. , 2007, Electroanalysis.

[5]  P. Piunno,et al.  Effects of Oligonucleotide Immobilization Density on Selectivity of Quantitative Transduction of Hybridization of Immobilized DNA , 2000 .

[6]  E. Wierzbiński,et al.  In situ electrochemical distance tunneling spectroscopy of ds-DNA molecules. , 2006, Langmuir : the ACS journal of surfaces and colloids.

[7]  U. Rant,et al.  Excessive counterion condensation on immobilized ssDNA in solutions of high ionic strength. , 2003, Biophysical journal.

[8]  F. Lisdat,et al.  The use of electrochemical impedance spectroscopy for biosensing , 2008, Analytical and bioanalytical chemistry.

[9]  I. Willner,et al.  Probing Biomolecular Interactions at Conductive and Semiconductive Surfaces by Impedance Spectroscopy: Routes to Impedimetric Immunosensors, DNA‐Sensors, and Enzyme Biosensors , 2003 .

[10]  J. C. Hoogvliet,et al.  Electrochemical pretreatment of polycrystalline gold electrodes to produce a reproducible surface roughness for self-assembly: a study in phosphate buffer pH 7.4 , 2000, Analytical chemistry.

[11]  Itamar Willner,et al.  Electronic transduction of biocatalytic transformations on nucleic acid-functionalized surfaces , 2001 .

[12]  R. Hintsche,et al.  Label-free impedance detection of oligonucleotide hybridisation on interdigitated ultramicroelectrodes using electrochemical redox probes. , 2005, Biosensors & bioelectronics.

[13]  J. Justin Gooding,et al.  DNA Biosensor Concepts Based on a Change in the DNA Persistence Length upon Hybridization , 2006 .

[14]  H. White,et al.  Voltammetry of molecular films containing acid/base groups , 1993 .

[15]  T. M. Herne,et al.  Characterization of DNA Probes Immobilized on Gold Surfaces , 1997 .

[16]  K. Niki,et al.  Double-Layer-Capacitance Titration of Self-Assembled Monolayers of ω-Functionalized Alkanethiols on Au(111) Surface , 2000 .

[17]  F. Illas,et al.  Stability of binary SAMs formed by omega-acid and alcohol functionalized thiol mixtures. , 2009, Langmuir : the ACS journal of surfaces and colloids.

[18]  Pedro Estrela,et al.  Optimization of DNA immobilization on gold electrodes for label-free detection by electrochemical impedance spectroscopy. , 2008, Biosensors & bioelectronics.

[19]  Keying Zhang,et al.  Fabrication of a Sensitive Impedance Biosensor of DNA Hybridization Based on Gold Nanoparticles Modified Gold Electrode , 2008 .

[20]  C. Bustamante,et al.  Ten years of tension: single-molecule DNA mechanics , 2003, Nature.

[21]  Yi-Tao Long,et al.  AC impedance spectroscopy of native DNA and M-DNA. , 2003, Biophysical journal.

[22]  Eileen M. Spain,et al.  Orienting DNA helices on gold using applied electric fields , 1998 .

[23]  W. Schuhmann,et al.  Label‐Free Detection of DNA Hybridization in Presence of Intercalators Using Electrochemical Impedance Spectroscopy , 2009 .

[24]  W. Olson,et al.  Approach to the limit of counterion condensation , 1990, Biopolymers.

[25]  Marc Tornow,et al.  Electrical manipulation of oligonucleotides grafted to charged surfaces. , 2006, Organic & biomolecular chemistry.

[26]  P. He,et al.  Impedance-Based DNA Biosensor Employing Molecular Beacon DNA as Probe and Thionine as Charge Neutralizer , 2006 .

[27]  S. Creager,et al.  Consequences of microscopic surface roughness for molecular self-assembly , 1992 .

[28]  Shaojun Dong,et al.  Kinetic study of DNA/DNA hybridization with electrochemical impedance spectroscopy , 2007 .

[29]  Stuart C. Burris,et al.  The Effect of Surface Preparation on Apparent Surface pKa's of ω-Mercaptocarboxylic Acid Self-Assembled Monolayers on Polycrystalline Gold , 2008 .

[30]  Marc Tornow,et al.  Structural properties of oligonucleotide monolayers on gold surfaces probed by fluorescence investigations. , 2004, Langmuir : the ACS journal of surfaces and colloids.

[31]  U. Rant,et al.  Dissimilar kinetic behavior of electrically manipulated single- and double-stranded DNA tethered to a gold surface. , 2006, Biophysical journal.

[32]  I. Willner,et al.  Electronic Transduction of Polymerase or Reverse Transcriptase Induced Replication Processes on Surfaces: Highly Sensitive and Specific Detection of Viral Genomes. , 2001, Angewandte Chemie.

[33]  S. Satija,et al.  Using Self-Assembly To Control the Structure of DNA Monolayers on Gold: A Neutron Reflectivity Study , 1998 .

[34]  Heinz-Bernhard Kraatz,et al.  Interaction of metal ions and DNA films on gold surfaces: an electrochemical impedance study. , 2009, The Analyst.

[35]  Fred Lisdat,et al.  A label-free DNA sensor based on impedance spectroscopy , 2008 .

[36]  Yi-Tao Long,et al.  Electrochemical detection of single-nucleotide mismatches: application of M-DNA. , 2004, Analytical chemistry.

[37]  Pedro Estrela,et al.  Optimization of label-free DNA detection with electrochemical impedance spectroscopy using PNA probes. , 2008, Biosensors & bioelectronics.

[38]  C. Buess-Herman,et al.  Electrochemical detection of the immobilization and hybridization of unlabeled linear and hairpin DNA on gold , 2005 .

[39]  A. Steel,et al.  Electrochemical quantitation of DNA immobilized on gold. , 1998, Analytical chemistry.

[40]  Marc Madou,et al.  DNA hybridization detection by label free versus impedance amplifying label with impedance spectroscopy , 2006 .

[41]  Itamar Willner,et al.  Amplified DNA detection by electrogenerated biochemiluminescence and by the catalyzed precipitation of an insoluble product on electrodes in the presence of the doxorubicin intercalator. , 2002, Angewandte Chemie.