Molecular Ecology Notes (2004) 4, 67 – 69 doi: 10.1046/j.1471-8286.2003.00571.x © 2004 Blackwell Publishing Ltd Blackwell Publishing, Ltd. PRIMER NOTE Isolation of 10 microsatellite markers for mongoose lemurs (Eulemur mongoz) J. PASTORINI,* P. FERNANDO,† D. J. MELNICK† and M. R. J. FORSTNER‡ *Department of Zoology, University of Cambridge, Downing Street, Cambridge CB2 3EJ, UK, †Center for Environmental Research and Conservation and Department of Ecology, Evolution, and Environmental Biology, Columbia University, 1200 Amsterdam Avenue, New York, NY 10027, USA, ‡ Department of Biology, Texas State University, 601 University Drive, San Marcos, TX 78666, USA Abstract We present here 10 new microsatellite markers for the mongoose lemur (Eulemur mongoz), nine of which were isolated from E. mongoz and one from Lemur catta. At least 60 individuals were genotyped for each of the 10 loci. Mendelian inheritance at each locus was tested by genotyping five captive families with known pedigrees. All loci were polymorphic and demon- strated simple Mendelian inheritance. The microsatellite markers presented here will be useful for genetic surveys of wild E. mongoz, to gain insights into the genetic background of the captive population, and to aid in the conservation of the species’ genetic diversity. Keywords: Eulemur, genetic markers, Lemuridae, lemurs, microsatellites, Primates Received 16 September 2003; revision received 5 November 2003; accepted 5 November 2003 The lemurs represent a prosimian group endemic to Madagascar. The mongoose lemur is one of five species in the genus Eulemur , which belongs to the family Lemuridae. E. mongoz occurs in three geographically isolated popu- lations; on two Comorian islands (Anjouan and Mohéli) and in the northwest of Madagascar (Tattersall 1982). It is one of only two lemur species occurring outside Madaga- scar, and was probably introduced to the Comores by humans (Mittermeier et al . 1994; Pastorini et al . 2003). The species is listed in Appendix 1 of CITES, and is protected by law in Madagascar and on the Comores (Harcourt & Thornback 1990). In the wild, mongoose lemurs live in small family groups, consisting of an adult pair and associ- ated offspring (Curtis & Zaramody 1998). Currently, there are more than 100 mongoose lemurs in captivity. While microsatellite markers have been developed for other lemur species, their application to E. mongoz has not been reported . The microsatellite loci described here will allow collection of genetic data that can guide management of captive E. mongoz (Bettinger 2000) and contribute to our under- standing of group dynamics and breeding systems in wild populations. Genomic DNA was extracted from an E. mongoz tissue sample using a standard phenol-chloroform extraction (Sambrook et al . 1989). The DNA was digested with the restriction enzyme Sau 3AI. After treatment with Calf Intestinal Phosphatase (New England Biolabs) the DNA fragments were ligated into Bam HI-digested LITMUS 29 phagemid vector (New England Biolabs) using T4 DNA Ligase (New England Biolabs). One Shot® Max Efficiency DH5 α TM -T1 R (Invitrogen) competent cells were used for the transformation. The transformation was plated on agar containing ampicillin and X-gal. Insert-bearing clones were grown in liquid LB and plated onto nylon membranes. In total, 5088 clones with inserts were screened for micro- satellites using four [ γ 32 P]-dATP labelled oligonucleotide probes (CA) 16 (TC) 16 (GC) 16 , and (AT) 16 . A total of 373 posit- ive clones were identified and were then PCR amplified and sequenced using universal primers to determine the repeat length and the flanking DNA sequences. Of the 373 positive clones, 109 contained a motif with four or more repeats. From these 109, primers were designed for 16 loci (Em1–Em16). Primer pairs were first used to amplify and sequence each locus to ensure it contained the expected repetitive motif. Microsatellite polymorphism was assessed on four distantly related individuals of E. mongoz . Three loci showed no variability. For the 13 polymorphic loci the PCR Correspondence: Jennifer Pastorini. Fax: + 44 –1223 33 6676; E-mail: jp357@cam.ac.uk