Enhanced primers for amplification of DNA barcodes from a broad range of marine metazoans

Lobo, Jorge; Costa, Pedro M.; Teixeira, Marcos A. L.; Ferreira, Maria S. G.; Costa, Maria Helena Ferrão Ribeiro da; Costa, Filipe Oliveira

http://hdl.handle.net/10362/31568

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Autores

Lobo, Jorge

Costa, Pedro M.

Teixeira, Marcos A. L.

Ferreira, Maria S. G.

Costa, Maria Helena Ferrão Ribeiro da

Costa, Filipe Oliveira

Resumo(s)

BACKGROUND: Building reference libraries of DNA barcodes is relatively straightforward when specifically designed primers are available to amplify the COI-5P region from a relatively narrow taxonomic group (e.g. single class or single order). DNA barcoding marine communities have been comparatively harder to accomplish due to the broad taxonomic diversity and lack of consistently efficient primers. Although some of the so-called "universal" primers have been relatively successful, they still fail to amplify COI-5P of many marine animal groups, while displaying random success even among species within each group. Here we propose a new pair of primers designed to enhance amplification of the COI-5P region in a wide range of marine organisms. RESULTS: Amplification tests conducted on a wide range of marine animal taxa, rendered possible the first--time sequencing of DNA barcodes from eight separated phyla (Annelida, Arthropoda, Chordata, Cnidaria, Echinodermata, Mollusca, Nemertea and Platyhelminthes), comprising a total of 14 classes, 28 orders, 57 families, 68 genus and 76 species. CONCLUSIONS: These primers demonstrated to be highly cost-effective, which is of key importance for DNA barcoding procedures, such as for building comprehensive DNA barcode libraries of marine communities, where the processing of a large numbers of specimens from a wide variety of marine taxa is compulsory.

Descrição

This work was supported by FEDER through POFC-COMPETE and by national funds from "Fundacao para a Ciencia e a Tecnologia (FCT)" in the scope of the grants FCOMP-01-0124-FEDER-015429. Filipe O Costa benefitted from the Marie Curie European Reintegration Grant PERG02-GA-2007-224890, provided by the European Commission. The authors would also like to thank Carolina Madeira and Monica Landi for the initial molecular experiments, Luisa Borges and Ronaldo Sousa for the support on collections, Catia Bartilotti (IPMA), Nuno Gomes (CBMA) for the offer of crustacean specimens and Artur Oliveira (CBMA) for the alga specimen.