TECHNICAL NOTE Characterization of microsatellite loci developed for the Mexican four-eyed octopus Octopus maya Oscar E. Jua ´rez • Carlos Rosas • Leticia Arena • Luis Enrı ´quez • Faustino Camarena • Niall McKeown • Paul W. Shaw Received: 8 March 2013 / Accepted: 15 March 2013 Ó Springer Science+Business Media Dordrecht 2013 Abstract Ten polymorphic microsatellite DNA loci were isolated from Mexican four-eyed octopus Octopus maya, by construction of an enriched genomic library. Genotyp- ing of 35 individuals from Sabancuy Campeche, Me ´xico, revealed variable levels of locus polymorphism with an average of 9.2 alleles. The observed and expected hetero- zygosity per locus ranged from 0.53 to 0.93 and from 0.48 to 0.87, respectively. No evidence of linkage disequilib- rium was detected between pairs of loci and genotype proportions at all loci conformed to Hardy–Weinberg equilibrium expectations. The microsatellite loci developed constitute a suite of genetic markers applicable to sus- tainable fishery management for O. maya. Keywords Octopus maya Á Polymorphic microsatellite Á Fishery management The endemic Octopus maya represents one of the most important fisheries in Yucatan peninsula, Mexico. The fishery has been managed as a single unit, but emerging evidence suggests that perhaps there are different units and that the resource could be overexploited (Jua ´rez et al. 2010). The high intrapopulation variability of microsatel- lite markers and their relatively ease of genotyping, makes them a powerful tool to discriminate cephalopod popula- tions, and informing sustainable fishery management (Shaw et al. 1999). The isolation and characterization of the first microsat- ellite loci for O. maya are reported here. Microsatellites were isolated an enriched partial genomic library by method outlined by Glenn and Schable (2005). The geno- mic DNA was extracted from O. maya muscle tissue fol- lowing phenol:chloroform (1:1) method, subsequently DNA was digested with restriction Rsa-I enzyme (New England Biolabs) producing fragments suitable for the ligation of oligonucleotide linkers specific to the protocol. Linker-ligated DNA was amplified by polymerase chain reaction (PCR). Enrichment was then performed by selective hybridisation of biotin-labelled repeat motif oli- gonucleotide probes to the PCR products. Hybridised complexes were captured using streptavidin-coated mag- netic beads (DYNAL). Microsatellite enriched eluates were thereafter PCR amplified and cloned into bacterial vectors using the TOPO-TA cloning kit (Invitrogen). Recombinant colonies were identified by inactivation of the B-galacto- sidase gene. Recombinant colonies were individually transferred into 50 lL of 10MmTris-HCL (pH 8.5) and incubated at 95 °C for 10 min to promote plasmid DNA release. One microlitre of each plasmid extract was sub- mitted to PCR involving the standard M13 forward and M13 reverse primers. The amplification reaction contained 19 buffer, 1.5 mM MgCL 2 , 0.2 mMdNTPs, 0.2 U of Taq O. E. Jua ´rez (&) Departamento de Biotecnologı ´a Marina, Centro de Investigacio ´n Cientı ´fica y de Educacio ´n Superior de Ensenada, Carretera Ensenada-Tijuana # 3918, Ensenada, Baja California, Me ´xico e-mail: ojuarez@cicese.edu.mx C. Rosas Á L. Arena Unidad Multidisciplinaria de Docencia e Investigacio ´n, Facultad de Ciencias, Universidad Nacional Auto ´noma de Me ´xico, Puerto de abrigo s/n, Sisal, Yucata ´n, Me ´xico L. Enrı ´quez Á F. Camarena Universidad Auto ´noma de Baja California, Km. 103 Carretera Tijuana-Ensenada, Ensenada, Baja California, Me ´xico N. McKeown Á P. W. Shaw Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, Penglais, Aberystwyth, Ceredigion SY23 3BF, United Kingdom 123 Conservation Genet Resour DOI 10.1007/s12686-013-9912-x Author's personal copy