Amplified rDNA Restriction Analysis (ARDRA) for Identification and Phylogenetic Placement of 16S-rDNA Clones Academic Article uri icon

abstract

  • Amplified ribosomal DNA restriction analysis (ARDRA), an extension of restriction fragment- length polymorphism (RFLP) analysis, is a tool that relies on DNA polymorphism to study microbial diversity [Deng et al., 2008]. In the ARDRA approach, the 16S rRNA gene is amplified by PCR, using either universal or genus-specific primer sets. The amplicon is digested by restriction endonucleases (REs), followed by separation of the fragments on high-percentage agarose or acrylamide gels. The emerging profiles are then used either to cluster the community into genotypic groups or for strain typing, both in accordance with the digested fragments' profile [Tiedje et al., 1999]. Classically, ARDRA has been employed on either isolated strains or clones (as part of a clone-screening step prior to sequencing), with the aim of providing a level of insight into the phylogeny of unique operational taxonomic …

publication date

  • January 1, 2011