Open Access Highly Accessed Open Badges Research

Comparative performance of the 16S rRNA gene in DNA barcoding of amphibians

Miguel Vences1*, Meike Thomas2, Arie van der Meijden3, Ylenia Chiari3 and David R Vieites4

Author Affiliations

1 Institute for Biodiversity and Ecosystem Dynamics, Zoological Museum, University of Amsterdam, Mauritskade 61, 1092 AD Amsterdam, The Netherlands

2 Institute for Genetics, Evolutionary Genetics, University of Cologne, Weyertal 121, 50931 Köln, Germany

3 Department of Biology (Evolutionary Biology), University of Konstanz, 78457 Konstanz, Germany

4 Department of Integrative Biology, Museum of Vertebrate Zoology, 3101 Valley Life Sciences Bldg., University of California, Berkeley, CA 94720-3160, USA

For all author emails, please log on.

Frontiers in Zoology 2005, 2:5  doi:10.1186/1742-9994-2-5

Published: 16 March 2005



Identifying species of organisms by short sequences of DNA has been in the center of ongoing discussions under the terms DNA barcoding or DNA taxonomy. A C-terminal fragment of the mitochondrial gene for cytochrome oxidase subunit I (COI) has been proposed as universal marker for this purpose among animals.


Herein we present experimental evidence that the mitochondrial 16S rRNA gene fulfills the requirements for a universal DNA barcoding marker in amphibians. In terms of universality of priming sites and identification of major vertebrate clades the studied 16S fragment is superior to COI. Amplification success was 100% for 16S in a subset of fresh and well-preserved samples of Madagascan frogs, while various combination of COI primers had lower success rates.COI priming sites showed high variability among amphibians both at the level of groups and closely related species, whereas 16S priming sites were highly conserved among vertebrates. Interspecific pairwise 16S divergences in a test group of Madagascan frogs were at a level suitable for assignment of larval stages to species (1–17%), with low degrees of pairwise haplotype divergence within populations (0–1%).


We strongly advocate the use of 16S rRNA as standard DNA barcoding marker for vertebrates to complement COI, especially if samples a priori could belong to various phylogenetically distant taxa and false negatives would constitute a major problem.