Molecular Ecology Resources (2008) 8, 1477–1479 doi: 10.1111/j.1755-0998.2008.02230.x © 2008 The Authors Journal compilation © 2008 Blackwell Publishing Ltd Blackwell Publishing Ltd PERMANENT GENETIC RESOURCES Isolation and characterization of microsatellite loci in Palinurus elephas F . PALERO and M . PASCUAL Departament de Genètica, Facultat de Biologia, Universitat de Barcelona, Av. Diagonal 645, 08028 Barcelona, Spain Abstract The European spiny lobster (Palinurus elephas) mean annual catches have decreased alarm- ingly during recent decades along its entire distribution area due to stock over-exploitation, which makes it a primary target for conservation plans. A total of 164 microsatellite loci were isolated from a genomic library of P. elephas enriched for CA, GA, CAA and GATA repeats. A total of 15 polymorphic loci have been screened in 48 individuals. High numbers of alleles per locus (averaging 20 ± 10.5) and observed heterozygosity (averaging 0.789 ± 0.197) have been detected. None of the pairs of loci showed significant linkage disequilibrium. Two of the loci (Pael1 and Pael2) showed significant departure from Hardy–Weinberg equilibrium in Sagres, while Pael38 showed significant departure in Tunis. These highly polymorphic markers will be useful in determining the spatial patterns of genetic diversity between and within populations of Palinurus elephas. Keywords: conservation genetics, crustacea, gene flow, microsatellites, Palinurus Received 17 February 2008; revision accepted 20 March 2008 The European spiny lobster (Palinurus elephas) is a palinurid lobster present in the Mediterranean Sea and along eastern Atlantic coasts from Morocco (28°N) to Norway (60°N). Mean annual catches of P. elephas have decreased alarmingly during recent decades along its entire distribution area due to stock over-exploitation (Goñi & Latrouite 2005). In order to establish proper management plans, it is crucial to define the population genetic structure of the endangered species. A plausible hypothesis would be that no population structuring is present in P. elephas, given the large dispersal potential of its larval phase (Palero & Abelló 2007). However, these dispersal capabilities could be counteracted by other factors. In fact, it has been shown that juveniles of the congeneric species Palinurus gilchristi migrate along distances of up to 700 km (Groeneveld & Branch 2002), which could modify the dispersal effect of the larval phase (Booth 1994). Furthermore, genetic differentiation has been detected among populations of this species, as revealed by the COI mtDNA gene, suggesting that complex oceanographic circulation patterns can affect population structure (Palero et al. 2008). Nevertheless, multiple markers should be analysed in order to unveil its population genetic structure. Herein, we report a set of microsatellite markers isolated from P. elephas, which will allow inferring the patterns of connectivity between populations and lead to the develop- ment of correct conservation plans. We constructed an enriched and partial genomic library following the FIASCO protocol (fast isolation by AFLP of sequences containing repeats, Zane et al. 2002). One pleopod sampled from a living individual from Galicia (Spain) and preserved in 100% ethanol was used for obtaining genomic DNA using the QIAamp DNA Mini Kit (QIAGEN Inc). Genomic DNA was simultaneously digested with MseI (New England BioLabs Inc.) and ligated to MseI-adapters (MseI-A, 5′-TACTCAGGACTCAT-3′; MseI-B, 5′-GACGAT- GAGGTCCTGAG-3′) for 3 hours. Enrichment was performed using the Streptavidin Magnesphere Paramagnetic Particles Kit (Promega) with four biotinylated probes [(CA) 15 (GA) 15 (CAA) 7 and (GATA) 7 ]. Recovered DNA was amplified via polymerase chain reaction (PCR) using MseI-N (5′-GATGAGTCCTGAGTAAN-3′) primers and sub- sequently cloned using the pGEM-T Easy Vector System II (Promega). Positive clones were detected using digoxigenin-end-labelled probes following the Estoup and Turgeon protocol (www.inapg.inra.fr/dsa/microsat /microsat.htm). Approximately 1500 colonies were screened for microsatellites yielding 247 positive clones, 138 of which Correspondence: Ferran Palero, Fax: 0034 934034420; E-mail: ferranpalero@ub.edu