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

Author:

Kris Richard Martin1,Felder Stephen1,Deyholos Michael1,Lambert Georgina M.1,Hinton James1,Botros Ihab1,Martel Ralph1,Seligmann Bruce1,Galbraith David W.1

Affiliation:

1. 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.)

Abstract

Abstract 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.

Publisher

Oxford University Press (OUP)

Subject

Plant Science,Genetics,Physiology

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