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Published 2009 | Published + Supplemental Material
Journal Article Open

Real-time DNA microarray analysis

Abstract

We present a quantification method for affinity-based DNA microarrays which is based on the real-time measurements of hybridization kinetics. This method, i.e. real-time DNA microarrays, enhances the detection dynamic range of conventional systems by being impervious to probe saturation in the capturing spots, washing artifacts, microarray spot-to-spot variations, and other signal amplitude-affecting non-idealities. We demonstrate in both theory and practice that the time-constant of target capturing in microarrays, similar to all affinity-based biosensors, is inversely proportional to the concentration of the target analyte, which we subsequently use as the fundamental parameter to estimate the concentration of the analytes. Furthermore, to empirically validate the capabilities of this method in practical applications, we present a FRET-based assay which enables the real-time detection in gene expression DNA microarrays.

Additional Information

© The Author(s) 2009. Published by Oxford University Press. Received May 18, 2009. Revised July 27, 2009. Accepted July 30, 2009. ACKNOWLEDGEMENTS The authors are grateful to Vijaya Kumar for experimental assistance with microarray manufacture and target labeling. They also want to thank Dr Scott Fraser for technical feedback in the imaging aspects of this project. FUNDING Grubstake grant from Caltech; Millard and Muriel Jacobs Genetics Laboratory at the California Institute of Technology; David and Lucille Packard Foundation. Funding for open access charge: University of Texas at Austin New Faculty Startup Fund. SUPPLEMENTARY DATA Supplementary Data are available at NAR Online.

Attached Files

Published - Hassibi2009p6447Nucleic_Acids_Res.pdf

Supplemental Material - nar-01026-met-k-2009-File003.doc

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