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PMID: 11872496 Published · ppublish English Evaluation Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

Quantitative detection of microbial genes by using DNA microarrays.

Applied and environmental microbiology ·Vol. 68 ·No. 3 ·2002-03-00 ·Pages 1425-30

Cho JC, Tiedje JM

Abstract

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.

MeSH Terms
Dioxygenases Gene Expression Profiling Gram-Negative Bacteria/enzymology,genetics,immunology Multienzyme Complexes/genetics Nitrate Reductase Nitrate Reductases/genetics O Antigens/genetics Oligonucleotide Array Sequence Analysis Oxygenases/genetics
Chemicals
Multienzyme Complexes O Antigens Oxygenases Dioxygenases naphthalene dioxygenase Nitrate Reductases Nitrate Reductase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Cho Jae-Chang
Center for Microbial Ecology, Plant and Soil Sciences Bldg., Michigan State University, East Lansing, MI 48824, USA.
Tiedje James M
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Article Info
Journal
Applied and environmental microbiology
Abbr.
Appl Environ Microbiol
ISSN
0099-2240
DOI
10.1128/AEM.68.3.1425-1430.2002
Published
2002-03-00
Pages
1425-30
Language
English
Region
United States
NLM ID
7605801
PMCID
PMC123775
Subset
IM
External Links
PubMed source DOI: 10.1128/AEM.68.3.1425-1430.2002 PMC Full Text View journal details
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