Quantitative detection of microbial genes by using DNA microarrays.
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.
Cho
Jae-Chang
JC
Center for Microbial Ecology, Plant and Soil Sciences Bldg., Michigan State University, East Lansing, MI 48824, USA.
Tiedje
James M
JM
D049308:Dioxygenases; D020869:Gene Expression Profiling; D006090:Gram-Negative Bacteria / Q000201:enzymology / Q000235:genetics* / Q000276:immunology; D009097:Multienzyme Complexes / Q000235:genetics; D050901:Nitrate Reductase; D009565:Nitrate Reductases / Q000235:genetics; D019081:O Antigens / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D010105:Oxygenases / Q000235:genetics
D009097:Multienzyme Complexes; D019081:O Antigens; D010105:Oxygenases; D049308:Dioxygenases; C038703:naphthalene dioxygenase; D009565:Nitrate Reductases; D050901:Nitrate Reductase