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Published on May 11, 2007; 10.1104/pp.107.098681

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Received February 28, 2007
Accepted May 5, 2007

High-Throughput, High-Sensitivity Analysis of Gene Expression in Arabidopsis

Richard Martin Kris *, Stephen Felder , Michael Deyholos , Georgina M. Lambert , James Hinton , Ihab Botros , Ralph Martel , Bruce Seligmann , and David W. Galbraith

NuvoGen Research, L.L.C., P.O. Box 64326, Tucson, Arizona 85728; High Throughput Genomics, Inc., 6296 E. Grant Road, Tucson, Arizona 85712; Department of Plant Sciences, University of Arizona, Tucson, Arizona 85721

* Corresponding author; email: rkris{at}nuvogenresearch.com.

High-throughput gene expression analysis of genes expressed during salt stress was performed using a novel multiplexed quantitative Nuclease Protection Assay that involves customized DNA microarrays printed within the individual wells of 96-well plates. The levels of expression of the transcripts from sixteen different genes were quantified within crude homogenates prepared from Arabidopsis thaliana plants also grown in a 96-well plate format. Examples are provided of the high degree of reproducibility of quantitative dose-response data, and of the sensitivity of detection of changes in gene expression within limiting amounts of tissue. The lack of requirement for RNA purification renders the assay particularly suited for high-throughput gene expression analysis, and for the discovery of novel chemical compounds that specifically modulate the expression of endogenous target genes.




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