16S rRNA gene-based analysis of microbial community by whole-genome amplification and minigel-single-strand conformation polymorphism technique.
We have developed an analytical technique for the 16S rRNA gene that comprises whole-genome amplification and the polymerase chain reaction (PCR)-minigel-single-strand conformation polymorphism technique (WGA-SSCP). Under optimal conditions, SSCP bands could be detected when genomic DNA from bacteria of interest comprised 0.5% or more of the specimen. This method will be effective for the identification of nonculturable bacteria in a microbial community.
Oto
Michiei
M
Department of Biotechnology, Tokyo Technical College, Higashi, Kunitachi-shi, Tokyo 186-0002, Japan. ooto@tera-house.ac.jp
Suda
Wataru
W
Shinoyama
Hirofumi
H
D004269:DNA, Bacterial / Q000032:analysis*; D004783:Environmental Microbiology; D016680:Genome, Bacterial; D016133:Polymerase Chain Reaction / Q000379:methods*; D018807:Polymorphism, Single-Stranded Conformational; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D012680:Sensitivity and Specificity
D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S