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Rapid Method for Coextraction of DNA and RNA from Natural Environments for Analysis of Ribosomal DNA- and rRNA-Based Microbial Community Composition

Applied and Environmental Microbiology · 2000 · Vol. 66(12) · pp. 5488–5491

Robert I. Griffiths✉(Newcastle University)Andrew S. Whiteley(Ecologie Microbienne Lyon)Anthony G. O’Donnell(Newcastle University)Mark Bailey(Ecologie Microbienne Lyon)

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Abstract

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.

Microbial Community Ecology and Physiology Genomics and Phylogenetic Studies Environmental DNA in Biodiversity StudiesRibosomal RNATemperature gradient gel electrophoresisBiologyRibosomal DNAMicrobial population biologyDNADNA extractionNucleic acid18S ribosomal RNA16S ribosomal RNA

MeSH terms

BacteriaDNA, BacterialDNA, RibosomalElectrophoresis, Agar GelEnvironmental MicrobiologyRNA, BacterialRNA, Ribosomal, 16SSoil MicrobiologyPolymerase Chain ReactionDNA FingerprintingEcosystem

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