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

作者：Friedrich von Wintzingerode, Ulf B. Göbel, Erko Stackebrandt · 发表于：FEMS Microbiology Reviews · 年份：1997 · DOI：10.1111/j.1574-6976.1997.tb00351.x · 被引用次数：2057 · 研究领域：Environmental DNA in Biodiversity Studies、Microbial Community Ecology and Physiology、Genomics and Phylogenetic Studies

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.