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

The conserved regions of tRNA(PHE) and 16S rRNA in the vertebrate mitochond rial genome were compared in order to design the primers, 12SR and 12SL. Th ese 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 li neages represented by the species listed in the text. There is little lengt h variation of the PCR product among different taxa. Further sequence analy sis revealed that the fragment contains complete lengths of 12S rRNA and tR NA(VAL), and that the length of 16S rRNA is 200 bp. In tests through ail re presentative taxa investigated, the above 2 primers could amplify the compl ete 12S rRNA gene from all representative taxa investigated. As the 12S rRN A 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.