Rapid Method for Coextraction of DNA and RNA from Natural Environments for Analysis of Ribosomal DNA- and rRNA-Based Microbial Community Composition
Быстрый метод совместной экстракции ДНК и РНК из природных сред для анализа состава микробных сообществ на основе рибосомной ДНК и рРНК
2000-12-01
SCID: 54.1/pr9kk2m5
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16S rRNA diversityDNA and RNA coextractiondenaturing gradient gel electrophoresisgrassland soil microbial communitiesreverse transcription-PCR
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Abstract (AI)
A rapid protocol for the extraction of total nucleic acids from environmental samples is described. The method facilitates concomitant assessment of microbial 16S rRNA diversity by PCR and reverse transcription-PCR amplification from a single extraction. Denaturing gradient gel electrophoresis microbial community analysis differentiated the active component (rRNA derived) from the total bacterial diversity (ribosomal DNA derived) down the horizons of an established grassland soil.
Key Findings
1
A rapid protocol extracts total nucleic acids from environmental samples for simultaneous DNA- and RNA-based microbial community analysis.
2
A single extraction supports PCR and reverse-transcription PCR assessment of microbial 16S rRNA diversity.
3
Denaturing gradient gel electrophoresis distinguished active microbial communities based on rRNA from total bacterial diversity based on ribosomal DNA.
4
The method differentiated active and total bacterial community composition across the horizons of an established grassland soil.
Research Object
Microbial communities in established grassland soil across soil horizons
Research Subject
The relationship between total bacterial diversity and the active microbial community component, assessed through ribosomal DNA- and rRNA-based 16S rRNA profiles
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2000-12-01
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