Quantitative detection of microbial genes by using DNA microarrays

被引：103

作者：

Cho, JC

Tiedje, JM

机构：

[1] Michigan State Univ, Ctr Microbial Ecol, E Lansing, MI 48824 USA

[2] Michigan State Univ, Dept Microbiol & Mol Genet, E Lansing, MI 48824 USA

[3] Hankuk Univ Foreign Studies, Dept Environm Sci, Yongin 449791, Kyoungki Do, South Korea

来源：

APPLIED AND ENVIRONMENTAL MICROBIOLOGY | 2002年 / 68卷 / 03期

关键词：

D O I：

10.1128/AEM.68.3.1425-1430.2002

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

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.

引用

页码：1425 / 1430

页数：6

共 23 条

[1] APPLICATIONS OF MAGNETIC BEADS WITH COVALENTLY ATTACHED OLIGONUCLEOTIDES IN HYBRIDIZATION - ISOLATION AND DETECTION OF SPECIFIC MEASLES-VIRUS MESSENGER-RNA FROM A CRUDE CELL LYSATE [J].