title: Increasing hybridization rate and sensitivity of DNA microarrays using isotachophoresis.
abstract: We present an on-chip electrokinetic method to increase the reaction kinetics and sensitivity of DNA microarray hybridization. We use isotachophoresis (ITP) to preconcentrate target molecules in solution and transport them over the immobilized probe sites of a microarray, greatly increasing the binding reaction rate. We show theoretically and experimentally that ITP-enhanced microarrays can be hybridized much faster and with higher sensitivity than conventional methods. We demonstrate our assay using a microfluidic system consisting of a PDMS microchannel superstructure bonded onto a glass slide on which 60 spots of 20-27 nt ssDNA oligonucleotide probes are immobilized. Our 30 min assay results in an 8.2 fold higher signal than the conventional overnight hybridization at 100 fM target concentration. We show rapid and quantitative detection over 4 orders of magnitude dynamic range of target concentration with no increase in the nonspecific signal. Our technique can be further multiplexed for higher density microarrays and extended for other reactions of target-surface immobilized ligands.
authors.affiliation: Department of Mechanical Engineering, Stanford University, CA 94305, USA.
authors.forename: Crystal M
authors.initials: CM
authors.lastname: Han
authors.forename: Evaldas
authors.initials: E
authors.lastname: Katilius
authors.forename: Juan G
authors.initials: JG
authors.lastname: Santiago
mesh_terms: D004277:DNA, Single-Stranded / Q000737:chemistry / Q000378:metabolism; D061925:Immobilized Nucleic Acids / Q000737:chemistry / Q000378:metabolism; D057625:Isotachophoresis / Q000379:methods*; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D008856:Microscopy, Fluorescence; D008956:Models, Chemical; D009693:Nucleic Acid Hybridization / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation / Q000379:methods*
chemical_list: D004277:DNA, Single-Stranded; D061925:Immobilized Nucleic Acids