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FEMS Microbiol Rev. 1997 Nov;21(3):213-29.

Determination of microbial diversity in environmental samples: pitfalls of PCR-based rRNA analysis.

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Institut für Mikrobiologie und Hygiene, Universitätsklinikum Charité, Berlin, Germany.


After nearly 10 years of PCR-based analysis of prokaryotic small-subunit ribosomal RNAs for ecological studies it seems necessary to summarize reported pitfalls of this approach which will most likely lead to an erroneous description on the microbial diversity of a given habitat. The following article will cover specific aspects of sample collection, cell lysis, nucleic acid extraction, PCR amplification, separation of amplified DNA, application of nucleic probes and data analysis.

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