TECHNICAL NOTE Microsatellite markers for the proboscis monkey (Nasalis larvatus) M. Salgado-Lynn • D. W. G. Stanton • R. Sakong • J. Cable • B. Goossens • M. W. Bruford Received: 21 July 2010 / Accepted: 21 July 2010 / Published online: 1 August 2010 Ó Springer Science+Business Media B.V. 2010 Abstract We describe eight polymorphic microsatellite loci for the proboscis monkey (Nasalis larvatus). These markers were tested with 33 samples, collected from Sabah and exhibited a mean of 6.25 alleles per locus and a mean expected heterozygosity of 0.674. All but one locus were in Hardy–Weinberg equilibrium, and no evidence for linkage disequilibrium was detected between any loci. Another 30 loci were isolated but remain to be fully examined. These markers should be useful for the future study of population genetic diversity and genetic structure in this emblematic species. Keywords Proboscis monkey Á Microsatellite loci Á Population genetics Á Nasalis larvatus Classified as endangered by IUCN (2010) and listed in Appendix I of CITES (UNEP-WCMC, 2010), the probos- cis monkey (Nasalis larvatus van Wurmb 1787) is endemic to the island of Borneo. Its distribution is restricted to lowland coastal and riverine forests, mangrove, and peat swamp. With a declining population, major threats include hunting, fire and, most importantly, anthropogenic habitat loss and fragmentation (Meijaard and Nijman 2000; Sha et al. 2008). Despite its uniqueness and conservation status, limited research on genetic variation in proboscis monkeys has been carried out due to a lack of reliable genetic markers and challenging sample collection (Jalil 2007). Here, we describe the isolation and characterization of microsatellite markers which can be used for individual and population-level genetic analyses, suitable for both invasive and non-invasive samples, for the conservation of this species. Muscle samples were opportunistically collected from two deceased proboscis monkeys at Lok Kawi Wildlife Park, and from two road killed animals, in Sabah, Malaysia; the former were used for the isolation of microsatellite loci. Fecal samples from another 29 wild individuals, also from Sabah, were collected as part of a population study and were used for characterizing the markers along with the tissue samples. Stool samples were stored in 70% ethanol, and muscle samples in a –70°C freezer. Faecal DNA was extracted via the DNA Stool Mini Kit (Qiagen GMBH, Germany) using a previously described protocol (Goossens et al. 2005). Tissue samples were extracted with DNeasy Blood & Tissue Kit (Qiagen GMBH, Germany) following the recommendations of the manufacturer, with minor modifications during elution (namely, 5 min incubation at 70°C with buffer AE, which was also preheated at the same temperature). To verify the existence of primate DNA from faecal extracts, a partial mitochondrial Control Region fragment was amplified using species specific primers (Jalil 2007) while DNA from the muscle samples was visualized in agarose gels (1.5%) and quantified by spectrofluorometry (Invitrogen’s Quant-iT TM PicoGreen Ò Kit microtiter assay, Molecular Devices’ SOFTmax Pro Ò ). Genomic libraries were constructed based on the pro- tocol from Glenn and Schable (2005). DNA was digested M. Salgado-Lynn (&) Á D. W. G. Stanton Á J. Cable Á B. Goossens Á M. W. Bruford School of Biosciences, Cardiff University, Museum Avenue, Cardiff CF10 3AX, UK e-mail: salgado-lynnm@cf.ac.uk R. Sakong Red Ape Encounters, Kinabatangan Orangutan Conservation Project, PO Box 3109, 90734 Sandakan, Sabah, Malaysia B. Goossens Danau Girang Field Centre, c/o Sabah Wildlife Department, Wisma Muis, 88100 Kota Kinabalu, Sabah, Malaysia 123 Conservation Genet Resour (2010) 2:159–163 DOI 10.1007/s12686-010-9295-1