An advanced molecular strategy to identify bacterial communities on art objects

Journal of Microbiological Methods
C Schabereiter-GurtnerS Rölleke

Abstract

The application of culture-independent techniques based on molecular biological methods, especially on the PCR amplification of 16S rRNA genes, attempts to overcome some shortcomings of conventional cultivation methods and reveals far more complex bacterial communities on art objects than can be shown by cultivation methods. One of the major challenges of investigating microbial growth on art objects by molecular means is the extraction of DNA, due to small sample amounts and PCR inhibitors. In the present study, we introduce a DNA extraction protocol, which allowed the extraction of PCR-amplifiable DNA from samples derived from lime wall paintings and loamy soil underground. The DNA extracts were used to amplify 16S ribosomal fragments, which were subsequently analyzed by denaturing gradient gel electrophoresis (DGGE). In parallel with the DGGE analysis, clone libraries containing PCR fragments of the ribosomal gene were constructed and clones were screened by DGGE. Clone libraries allow the inclusion of the entire 16S rDNA sequence in the phylogenetic analyses of microorganisms, providing a more reliable phylogenetic identification of microorganisms than is obtained from sequence analyses of excised and directly sequenced DGG...Continue Reading

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Citations

Jan 5, 2006·Microbial Ecology·Christopher J McNamaraRalph Mitchell
Oct 18, 2006·Applied Microbiology and Biotechnology·P Fernandes
Oct 9, 2013·Applied Microbiology and Biotechnology·Katja Sterflinger, Guadalupe Piñar
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Feb 3, 2009·Journal of Environmental Science and Health. Part A, Toxic/hazardous Substances & Environmental Engineering·Jee-Eun MinJae-Woo Park

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