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Journal of Bioscience and Bioengineering, Vol.102, No.5, 482-484, 2006
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.
Keywords:whole-genome amplification (WGA);polymerase chain reaction (PCR);single-strand conformation polymorphism (SSCP);16S rRNA gene;nonculturable bacteria