Direct profiling of environmental microbial populations by thermal dissociation analysis of native rRNAs hybridized to oligonucleotide microarrays

被引:69
作者
El Fantroussi, S
Urakawa, H
Bernhard, AE
Kelly, JJ
Noble, PA
Smidt, H
Yershov, GM
Stahl, DA
机构
[1] Univ Washington, Dept Civil & Environm Engn, Seattle, WA 98195 USA
[2] Argonne Natl Lab, Biochip Technol Ctr, Argonne, IL 60439 USA
[3] Loyola Univ, Dept Biol, Chicago, IL 60626 USA
关键词
D O I
10.1128/AEM.69.4.2377-2382.2003
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
Oligonucleotide microarrays were used to profile directly extracted rRNA from environmental microbial populations without PCR amplification. In our initial inspection of two distinct estuarine study sites, the hybridization patterns were reproducible and varied between estuarine sediments of differing salinities. The determination of a thermal dissociation curve (i.e., melting profile) for each probe-target duplex provided information on hybridization specificity, which is essential for confirming adequate discrimination between target and nontarget sequences.
引用
收藏
页码:2377 / 2382
页数:6
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