DNA_primers_for_amplification_of_mitochondrial_cytochrome.pdf

Molecular Marine Biology and Biotechnology (1994) 3(5), 294-299 DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates 0. Folmer, M. Black, W. Hoeh,* R. Lutz, andR. Vrijenhoek+ Center for Theoretical and Applied Genetics, and Institute of Marine and Coastal Science, Rutgers University, New Brunswick, New Jersey 08903-231 Abstract We describe universal DNA primers for polymer- ase chain reaction (PCR) amplification of a 710-bp fragment of the mitochondrial cytochrome c oxidasesubunit I gene (COI) from 11 invertebrate phyla:Echinodermata, Mollusca, Annelida, Pogonophora, Arthropoda, Nemertinea, Echiura, Sipuncula, Pla- tyhelminthes, Tardigrada, and Coelenterata, as well as the putative phylum Vestimentifera. Preliminary comparisons revealed that these COI primers gener- ate informative sequences for phylogenetic analyses at the species and higher taxonomic levels. Introduction The purpose of this short communication is to de- scribe universal DNA primers for the polymerase chain reaction (PCR) amplification of a 710-bp frag- ment of the mitochondrial cytochrome c oxidase subunit I gene ( COI). This study was motivated by the recent discoveries of more than 230 new inver- tebrate species, comprising new genera, families, classes, orders, and potentially a new phylum, fromdeep-sea hydrothermal vent and cold-water sulfide or methane seep communities (Tunnicliffe, 1991). Our goal was to develop molecular techniques for phylogenetic studies of these diverse organisms. We focused on the mitochondrial cytochrome c oxi-dase subunit I (COI) gene because it appears to be among the most conservative protein-coding genesin the mitochondrial genome of animals (Brown, 1985), which was preferable for the evolutionary *Present address: Department of Biology, Dalhousie Univer- sity, Halifax, Nova Scotia, Canada. +Correspondence should be sent to this author. Copyright ? 1994 Blackwell Science, Inc. 294 time depths likely to be