Use of order-specific primers to investigate the methanogenic diversity in acetate enrichment system
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- Use of order-specific primers to investigate the methanogenic diversity in acetate enrichment system
- Shin, Seung Gu; Lee, Changsoo; Hwang, Kwanghyun; Ahn, Johng-Hwa; Hwang, Seokhwan
- Denaturing gradient gel electrophoresis; Diversity of methanogens; Order-specific primers; Real-time quantitative PCR; Taxonomic level
- Issue Date
- SPRINGER HEIDELBERG
- JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY, v.35, no.11, pp.1345 - 1352
- The applicability of order-specific primers in minimizing the possible underestimation of microbial diversity was evaluated via denaturing gradient gel electrophoresis (DGGE) analysis of a lab-scale anaerobic digester. Initially, a population analysis with real-time quantitative PCR demonstrated the existence of three methanogenic orders-Methanobacteriales, Methanomicrobiales, and Methanosarcinales-throughout the reaction period. DGGE analyses with three pairs of order-specific primers yielded eight operational taxonomic units (OTUs), whereas DGGE analysis with two independent Archaea-specific primers identified only five. Moreover, the order-specific primers amplified at least one OTU affiliated with each order, whereas no members of Methanobacteriales or Methanomicrobiales were identified with Archaea-specific primers in most samples. These findings provide evidence that order-specific analysis can detect the diversity of methanogens in greater detail than conventional Archaea-specific analysis.
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