Charging behavior of single-stranded DNA polyelectrolyte brushes.

DNA monolayers are widely used in fundamental and applied genomics and are versatile experimental models for elucidating the behavior of charged polymers at interfaces. The physical behavior of these systems is to a large extent governed by their internal ionic microenvironment, which is investigated here for layers of end-tethered, single-stranded DNA oligonucleotides (DNA brushes). Retention of counterions by the DNA brush manifests as lowered susceptibility of the interfacial capacitance to external salt conditions. A physical model based on concepts adapted from polymer science was used to further elucidate the connection between monolayer organization and its charging behavior. The data indicate a reorganization of the monolayer with changes in ionic strength and strand coverage that is consistent with that expected for a polyelectrolyte brush. A method for electrochemical quantification of strand coverage, based on shift of reduction potential for redox counterions associated with the DNA monolayer, is also described. These results provide guidance for development of label-free electrochemical diagnostics employing DNA monolayers and formulate a description of monolayer behavior within a polymer science framework.

Sensitive, label-free DNA diagnostics based on near-field microwave imaging.

Biological assays often rely on "reporter labels" to enhance measurement sensitivity, for example, by incorporation of a fluorescent dye or a nanoparticle into a nucleic acid or a protein. Use of labels, however, complicates sample preparation, increases assay costs, and can cause experimental artifacts by interfering with assay thermodynamics or limiting label stability. We evaluate near-field microwave imaging (NFMI) as an alternative, label-free technique for molecular diagnostics. Using DNA monolayers as an experimental model, NFMI is demonstrated to achieve sensitivities comparable to conventional fluorescence bioassays. Moreover, NFMI is shown to be compatible with imaging at resolutions required by microarray applications, as demonstrated by monitoring DNA hybridization in an array format.

Polymer-anchored DNA gene monolayers.

Monolayers of DNA chains of polymeric dimensions, considered here to be longer than approximately 100 nucleotides, are widely encountered in biomolecular diagnostics as well as present for a model system for investigating behavior of polyelectrolytes at interfaces. A major challenge in advancing such applications is assembling the DNA on the surface in a controlled way. Although covalent immobilization is expected to produce optimal stability, the multitude of potential reactive sites along the contour of long DNA molecules requires that any chemical transformations be strictly site-specific to preserve control over attachment geometry and function. A synthetic approach to fabricating monolayers of DNA genes on gold using polymeric anchor (adhesion) films is presented that (i) possesses stringent site-specificity of surface-attachment, (ii) exhibits excellent stability to elevated temperatures, allowing denaturation of duplex chains at 90 degrees C without loss of surface-linked strands, and (iii) achieves surface coverages suitable for investigating multichain polyelectrolyte behavior in regimes of strong interchain interactions.