FEMS Microbiology Ecology · 2002 · 15 citations · 23 references
Life SciencesMicrobiologyMolecular BiologyDiagnostic MicrobiologyMicrobial EcologyMicrobial Genetics16S Ribosomal Ribonucleic Acid AnalysisDegraded Dna AnalysisGenotypic Characterization16S Ribosomal Ribonucleic Acid Gene AnalysisGenomic Dna AnalysisPlasmid Vector ConstructionPlasmid Vector DesignPlasmid Vector EngineeringPolymerase Chain Reaction (Pcr) DiagnosticsPolymerase Chain Reaction (Pcr)-based IdentificationRestriction Enzyme Digestion
Abstract We report the presence of Actinomycetes in degraded sandstone monuments, and on examination of 173 samples we identified Nocardia restricta as particularly prevalent. In our procedure, the extracted bacterial DNA was the template in polymerase chain reaction (PCR) experiments in order to amplify specific regions of the 16S rDNA. The fidelity of amplified fragment was confirmed by nested-PCR or restriction enzyme specific cutting. To confirm the specificity of the assay, the amplified fragments were cloned in a convenient plasmid vector, the sequence analysed and compared with the expected DNA genomic portion.
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PCR protocols — A guide to methods and applications
Trends in Genetics · 1990 · 11.9K citations
16S ribosomal DNA amplification for phylogenetic study
W G Weisburg, Susan M. Barns, Dale A. Pelletier et al. · Journal of Bacteriology · 1991 · 11.8K citations · Full text
DNA recovery from soils of diverse composition
Jizhong Zhou, Mary Ann Bruns, J.M. Tiedje · Applied and Environmental Microbiology · 1996 · 2.9K citations · Full text