Universal primers for amplification of the complete mitochondrial 12S rRNA gene in vertebrates

The conserved regions of tRNA^PHE and 16S rRNA in the vertebrate mitochondrial genome were compared in order to design the primers, 12SR and 12SL. These universal primers can be broadly used to amplify a 1.3-kb DNA fragment by polymerase chain reaction (PCR) over a wide range of major vertebrate lineages represented by the species listed in the text. There is little length variation of the PCR product among different taxa. Further sequence analysis revealed that the fragment contains complete lengths of 12S rRNA and tRNA^VAL, and that the length of 16S rRNA is 200 bp. In tests through all representative taxa investigated, the above 2 primers could amplify the complete 12S rRNA gene from all representative taxa investigated. As the 12S rRNA gene is widely used for phylogenetic analyses among different hierarchies, the use of these primer sets for study of higher-category phylogenies in vertebrates now becomes possible.