Isolation and characterization of RNA from low-biomass deep-subsurface sediments
- 1 February 1995
- journal article
- research article
- Published by American Society for Microbiology in Applied and Environmental Microbiology
- Vol. 61 (2) , 763-768
- https://doi.org/10.1128/aem.61.2.763-768.1995
Abstract
Three methods for the isolation of microbial RNA from low-biomass deep-subsurface sediments have been developed and evaluated. RNA was isolated from samples taken from depths ranging from 173 to 217 m, and samples represented a variety of lithologies, including lacustrine, fluvial sand, and paleosol sediments. Cell numbers in these samples were estimated to be between log 4.0 and log 5.1/g on the basis of phospholipid fatty acid analysis. The most efficient method examined is based on the direct lysis of microbial cells followed by the extraction of RNA with alkaline phosphate buffers and subsequent inactivation of nucleases by extraction with guanidinium isothiocyanate. Estimated recoveries of mRNA for this method are approximately 26%. The recovered RNA included both mRNA and rRNA, as evidenced by the detection of sequences homologous to transcripts from the toluene-4-monooxygenase gene of Pseudomonas mendocina KR1 and bacterial, archaeal, and eukaryotic rRNA. An unexpectedly high relative concentration of archaeal rRNA (22 to 40%) was observed for these samples.Keywords
This publication has 13 references indexed in Scilit:
- Estimating the Growth Rate of Slowly Growing Marine Bacteria from RNA ContentApplied and Environmental Microbiology, 1993
- Direct extraction and purification of rRNA for ecological studiesApplied and Environmental Microbiology, 1993
- Archaea in coastal marine environments.Proceedings of the National Academy of Sciences, 1992
- Novel major archaebacterial group from marine planktonNature, 1992
- Cloning and characterization of a Pseudomonas mendocina KR1 gene cluster encoding toluene-4-monooxygenaseJournal of Bacteriology, 1991
- In Situ Biodegradation: Microbiological Patterns in a Contaminated AquiferScience, 1991
- Rapid Method for Direct Extraction of mRNA from Seeded SoilsApplied and Environmental Microbiology, 1991
- Genetic diversity in Sargasso Sea bacterioplanktonNature, 1990
- Phylogenetic Stains: Ribosomal RNA-Based Probes for the Identification of Single CellsScience, 1989
- Microbial Ecology of the Terrestrial SubsurfaceAdvances in applied microbiology, 1988