RDNA Amplification: Application of 16S rDNA-Based Methods for Bacterial Identification

Abstract

Continuing improvements in PCR and automated sequencing technologies, resulting in rapid acquisition of sequence data, have allowed these methods to be applied to the task of bacterial identification. The sequence of the 16S rDNA/RNA of bacterial strains has been demonstrated to be useful for phylogenetic analysis of different taxa. Comparative analysis of the partial 16S rDNA sequences from unidentified bacterial isolates with the extensive database now available can be used to assign the isolate to a genus or species within higher taxa of the domains Bacteria and Archaea. For isolates that are shown by partial 16S rDNA sequence analysis to be unique, the complete gene should be sequenced to determine more precisely the phylogenetic position of the isolates within the established taxonomic structure. The procedure involves the isolation of genomic DNA, the PCR-mediated amplification of the 16S rDNA, the purification and direct sequencing of the PCR products and the comparative analysis of 16S rDNA sequence data (Rainey et al., 1996).