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Issue 16, 2014
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Increasing hybridization rate and sensitivity of DNA microarrays using isotachophoresis

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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.

Graphical abstract: Increasing hybridization rate and sensitivity of DNA microarrays using isotachophoresis

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Publication details

The article was received on 26 Mar 2014, accepted on 20 May 2014 and first published on 20 May 2014


Article type: Paper
DOI: 10.1039/C4LC00374H
Author version available: Download Author version (PDF)
Citation: Lab Chip, 2014,14, 2958-2967
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    Increasing hybridization rate and sensitivity of DNA microarrays using isotachophoresis

    C. M. Han, E. Katilius and J. G. Santiago, Lab Chip, 2014, 14, 2958
    DOI: 10.1039/C4LC00374H

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