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Proceedings Paper

Multiplexed chemiluminescent assays in ArrayPlates for high-throughput measurement of gene expression
Author(s): Ralph R. Martel; Matthew P. Rounseville; Ihab W. Botros; Bruce E. Seligmann
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Paper Abstract

Multiplexed Molecular Profiling (MMP) assays for drug discovery are performed in ArrayPlates. ArrayPlates are 96- well microtiter plates that contain a 16-element array at the bottom of each well. Each element within an array measures one analyte in a sample. A CCD imager records the quantitative chemiluminescent readout of all 1,536 elements in a 96-well plate simultaneously. Since array elements are reagent modifiable by the end-user, ArrayPlates can be adapted to a broad range of nucleic acid- and protein-based assays. Such multiplexed assays are rapidly established, flexible, robust, automation-friendly and cost-effective. Nucleic acid assays in ArrayPlates can detect DNA and RNA, including SNPs and ESTs. A multiplexed mRNA assay to measure the expression of 16 genes is described. The assay combines a homogeneous nuclease protection assay with subsequent probe immobilization to the array by means of a sandwich hybridization followed with chemiluminescent detection. This assay was used to examine cells grown and treated in microplates and avoided cloning, transfection, RNA insolation, reverse transcription, amplification and fluorochrome labeling. Standard deviations for the measurement of 16 genes ranged from 3 percent to 13 percent in samples of 30,000 cells. Such ArrayPlates transcription assays are useful in drug discovery and development for target validation, screening, lead optimization, metabolism and toxicity profiling. Chemiluminescent detection provides ArrayPlates assays with high signal-to-noise readout and simplifies imager requirements. Imaging a 2D surface that contains arrays simplifies lens requirements relative to imaging columns of liquid in microtiter plate wells. The Omix imager for ArrayPlates is described.

Paper Details

Date Published: 21 June 2002
PDF: 9 pages
Proc. SPIE 4626, Biomedical Nanotechnology Architectures and Applications, (21 June 2002); doi: 10.1117/12.472069
Show Author Affiliations
Ralph R. Martel, High Throughput Genomics, Inc. (United States)
Matthew P. Rounseville, High Throughput Genomics, Inc. (United States)
Ihab W. Botros, High Throughput Genomics, Inc. (United States)
Bruce E. Seligmann, High Throughput Genomics, Inc. (United States)

Published in SPIE Proceedings Vol. 4626:
Biomedical Nanotechnology Architectures and Applications
Raymond P. Mariella; Catherine J. Murphy; Michelle Palmer; David A. Dunn; Darryl J. Bornhop; David A. Dunn; Raymond P. Mariella; Catherine J. Murphy; Dan V. Nicolau; Shuming Nie; Michelle Palmer; Ramesh Raghavachari; Darryl J. Bornhop; Ramesh Raghavachari; Shuming Nie; Ramesh Raghavachari, Editor(s)

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