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Applied and Environmental Microbiology, March 2002, p. 1425-1430, Vol. 68, No. 3
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.3.1425-1430.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Quantitative Detection of Microbial Genes by Using DNA Microarrays

Jae-Chang Cho1,2 and James M. Tiedje1,3*

Center for Microbial Ecology,1 Department of Microbiology and Molecular Genetics, Michigan State University, East Lansing, Michigan 48824,3 Department of Environmental Science, Hankuk University of Foreign Studies, Yongin Shi, Kyoungki Do, Korea 449-7912

Received 14 September 2001/ Accepted 17 December 2001

To quantify target genes in biological samples using DNA microarrays, we employed reference DNA to normalize variations in spot size and hybridization. This method was tested using nitrate reductase (nirS), naphthalene dioxygenase (nahA), and Escherichia coli O157 O-antigen biosynthesis genes as model genes and lambda DNA as the reference DNA. We observed a good linearity between the log signal ratio and log DNA concentration ratio at DNA concentrations above the method's detection limit, which was approximately 10 pg. This approach for designing quantitative microarrays and the inferred equation from this study provide a simple and convenient way to estimate the target gene concentration from the hybridization signal ratio.

* Corresponding author. Mailing address: Center for Microbial Ecology, Plant and Soil Sciences Bldg., Michigan State University, East Lansing, MI 48824. Phone: (517) 353-9021. Fax: (517) 353-2917. E-mail: tiedjej{at}msu.edu.

Applied and Environmental Microbiology, March 2002, p. 1425-1430, Vol. 68, No. 3
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.3.1425-1430.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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