Evaluation of a microplate DNA - DNA hybridization method compared with the initial renaturation method

Abstract

Previously reported hybridization data, obtained with the initial renaturation method (De Ley et al. 1970), were compared with data obtained with a microplate DNA - DNA hybridization method (Ezaki et al. 1989). The comparison was done for more than 82 hybridizations, within 4 sets of Bacillus/Virgibacillus, Myroides, Vibrio, and Xanthomonas/Stenotrophomonas strains, comprising DNA-base compositions ranging from 34 to 65%. Under the experimental conditions used, both methods were in very good correlation. With the microplate method, the variation between reciprocal hybridizations and repeated experiments was around 7%, while for the initial renaturation method, a 5% variation was calculated. It is concluded that the microplate method can be used as a reliable taxonomic tool.Key words: DNA, DNA hybridization, microplate method, initial renaturation method.