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Evaluation of DNA Extraction Methods for Molecular Analyses of Microbial Communities in Modern Calcareous Microbialites

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Ferran Garcia-Pichel, and Brian D Wade, Evaluation of DNA Extraction Methods for Molecular Analyses of Microbial Communities in Modern Calcareous Microbialites: .

Summary

We evaluated and optimized three rapid methods for extraction of high-quality DNA from carbonate-encrusted microbial communities using modern calcifying oncolites built by cyanobacteria and diatoms in a high-calcium freshwater river. Pulverization, acid (HCl) dissolution, and chelator-mediated (EDTA) dissolution of the carbonate matrix were used and optimized to liberate microbial cells from their mineral encasing. This was followed by cell lysis and DNA extraction and isolation. HCl dissolution yielded no measurable DNA, probably due to hydrolysis, whereas pulverization and EDTA dissolution yielded averages of 3.5 and 7.8 μg per gram of sample, respectively, of high molecular weight DNA. These DNA isolates could be used successfully [...]

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Title Citation Evaluation of DNA Extraction Methods for Molecular Analyses of Microbial Communities in Modern Calcareous Microbialites

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