PCR primers and amplification methods for 12S ribosomal DNA, the control region, cytochrome oxidase I, and cytochrome b in bufonids and other frogs, and an overview of PCR primers which have amplified DNA in amphibians successfully.

ABSTRACT

New primers (N = 24) for the amplification and sequencing of the complete or near complete 12S ribosomal DNA, about 1000 bp of the control region, 390 bp of cytochrome oxidase I, and the near complete cytochrome b are described. The 12S ribosomal DNA primers successfully amplify DNA in tetrapods; other primers successfully amplify DNA in bufonoids and other anurans. An overview of published literature and sequence data banks identified 170 mitochondrial and 96 nuclear DNA primers that have been used or are highly likely to be useful in amphibians. Primer sequences, their locations within genes, and sequence location and identity in Xenopus and human and/or mouse are presented for each primer. The utility of each primer was estimated by identifying the smallest, yet most inclusive, taxonomic category within which each primer has been successful. Primers from all published sources are mapped together. We hope that these new primers, as well as the list of primers that have been useful in amphibians, will encourage further systematic and population genetic studies of amphibians.