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BREAKTHROUGH TECHNOLOGIES

High-Throughput, High-Sensitivity Analysis of Gene Expression in Arabidopsis^1,[W],[OA]

NuvoGen Research, L.L.C., Tucson, Arizona 85728 (R.M.K., S.F.); Department of Plant Sciences, University of Arizona, Tucson, Arizona 85721 (M.D., G.M.L., D.W.G.); and High Throughput Genomics, Inc., Tucson, Arizona 85712 (J.H., I.B., R.M., B.S.)

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 16 different genes were quantified within crude homogenates prepared from Arabidopsis (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.

² Present address: Department of Biological Sciences, CW 405, Biological Sciences Centre, University of Alberta, Edmonton, Alberta, Canada T6G 2E9.

The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Richard Martin Kris (rkris{at}nuvogenresearch.com).

^[W] The online version of this article contains Web-only data.

^[OA] Open Access articles can be viewed online without a subscription.

www.plantphysiol.org/cgi/doi/10.1104/pp.107.098681

^* Corresponding author; e-mail rkris{at}nuvogenresearch.com; fax 520–232–9429.

Received February 28, 2007; accepted May 5, 2007; published May 11, 2007.

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