FRET-Based Real-Time DNA Microarrays
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, washing artifacts, microarray spot-to-spot variations, and other intensity-affecting impediments. We demonstrate in both theory and practice that the time-constant of target capturing is inversely proportional to the concentration of the target analyte, which we take advantage of as the fundamental parameter to estimate the concentration of the analytes. Furthermore, to experimentally 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.
Key wordsMicroarray Real-time Gene expression Time-constant FRET
We are grateful to Vijaya Kumar for experimental assistance with microarray manufacture and target labeling. We also want to thank Professor Scott Fraser at Caltech for technical feedback in the imaging aspects of this project.
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