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Applied and Environmental Microbiology, February 2003, p. 1214-1219, Vol. 69, No. 2
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.2.1214-1219.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Development of Sinorhizobium meliloti Pilot Macroarrays for Transcriptome Analysis

Hélène Bergès, Emmanuelle Lauber,* Carine Liebe, Jacques Batut, Daniel Kahn, Frans J. de Bruijn, and Frédéric Ampe

Laboratoire de Biologie Moléculaire des Relations Plantes-Microorganismes, UMR215 CNRS-INRA, 31326 Castanet-Tolosan Cedex, France

Received 8 July 2002/ Accepted 2 November 2002

In order to prepare for whole-genome expression analysis in Sinorhizobium meliloti, pilot DNA macroarrays were designed for 34 genes of known regulation. The experimental parameters assessed were the length of the PCR products, the influence of a tag at the 5' end of the primers, and the method of RNA labeling. Variance and principal-component analysis showed that the most important nonbiological parameter was the labeling method. The sizes of PCR products were also found to be important, whereas the influence of 5' tags was minimal. The variability between replicated spots on a membrane was found to be low. These experimental procedures were validated by analyzing the effects of microaerobic conditions on gene expression.

* Corresponding author. Mailing address: Chemin de Borde Rouge BP27, 31326 Castanet-Tolosan Cedex, France. Phone: 33 561 28 50 48. Fax: 33 561 28 50 61. E-mail: elauber{at}toulouse.inra.fr.

Applied and Environmental Microbiology, February 2003, p. 1214-1219, Vol. 69, No. 2
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.2.1214-1219.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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