New primers for the amplification and sequencing of nuclear loci in a taxonomically wide set of reptiles and amphibians
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We report new primers for the amplification and sequencing of 11 nuclear markers in squamate reptiles and anuran amphibians (five in squamates, six in anurans). Ten out of the 11 loci are introns (three of which are linked) that were amplified using an exon-primed, intron-crossing (EPIC) PCR strategy, whereas an eleventh locus spans part of a protein-coding gene. Squamate and anuran primers were initially developed for Lacerta schreiberi (Squamata: Lacertidae) and Pelodytes spp. (Anura: Pelodytidae), respectively. Cross-species amplification of the squamate markers was evaluated in four genera representing two additional families, whereas for anurans three genera corresponding to three additional families were tested. Three out of the five loci were successfully sequenced in all squamate taxa tested. Cross-amplification of the six anuran markers had lower, but still significant, success. We predict these markers will be of great utility for both population genetics and phylogenetic studies.
KeywordsAnura Squamata Nuclear sequence markers Introns
This work was partially financed by Fundação para a Ciência e a Tecnologia (FCT) research projects POCI/BIA-BDE/60911/2004, PTDC/BIA-BDE/69769/2006 and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) and FCT joint project CAPES/FCT 244/09. CP, HG and FS are supported by post-doctoral fellowships SFRH/BPD/28869/2006, SFRH/BPD/26555/2006 and SFRH/BPD/27134/2006, respectively and SR is supported by pre-doctoral grant SFRH/BD/17541/2004, all from FCT. BMC and TOB are supported by the research grants (BI/PTDC/BIA-BDE/69769/2006 and BI/PTDC/BIA-QOR/71492/2006, respectively). MV is supported by a post-doctoral grant CAPES 1362-07-0. CFBH thanks Fundação de Amparo à Pesquisa do Estado de S. Paulo and Conselho Nacional de Desenvolvimento Científico e Tecnológico for financial support. We further thank Raquel Godinho and Miguel Tejedo for providing tissue samples.
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