DETERMINATION OF MICROBIAL DIVERSITY IN ENVIRONMENTAL-SAMPLES - PITFALLS OF PCR-BASED RIBOSOMAL-RNA ANALYSIS

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