Isolation and characterization of nine microsatellite loci in a Malagasy endemic rodent, Eliurus carletoni (Rodentia: Ne
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TECHNICAL NOTE
Isolation and characterization of nine microsatellite loci in a Malagasy endemic rodent, Eliurus carletoni (Rodentia: Nesomyinae) Jean-Eric Rakotoarisoa • Carolyn A. Bailey Paula H. Hinger • Rick A. Brenneman • Edward E. Louis Jr.
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Received: 24 August 2012 / Accepted: 27 August 2012 / Published online: 14 September 2012 Ó Springer Science+Business Media B.V. 2012
Abstract We developed microsatellite markers for the recently described forest rodent, Eliurus carletoni, from an enriched genomic library. Nine loci composed of four dinucleotide, one trinucleotide, one tetranucleotide and three compound repeats were isolated and characterized using two wild populations. One locus was found to be monomorphic. For the polymorphic loci, the average number of alleles per locus was 7.13 and 8.38 for each population. Mean expected and observed heterozygosities were high (i.e. 0.76 and 0.80). Tests for linkage disequilibrium were not significant across all locus pairs. One locus tested significant for null alleles, but only one population exhibited a significant deviation from the Hardy– Weinberg Equilibrium (HWE) at this locus. All remaining loci show no evidence of departure from HWE. Overall, we identified eight polymorphic loci that may be used in conservation and population genetics studies of E. carletoni. Keywords Microsatellites Eliurus Mammals Madagascar
J.-E. Rakotoarisoa Ecology and Evolutionary Biology Department, Yale University, 21 Sachem St., New Haven, CT 06511, USA J.-E. Rakotoarisoa (&) School of Biological Sciences, Illinois State University, 125 South Fell Ave, Normal, IL 61790, USA e-mail: [email protected] C. A. Bailey P. H. Hinger R. A. Brenneman E. E. Louis Jr. Grewcock Center for Conservation and Research, Omaha’s Henry Doorly Zoo, 3701 South 10th St., Omaha, NE 68107, USA e-mail: [email protected]
Introduction The recently described forest rodent, Eliurus carletoni (Rodentia: Nesomyinae), is restricted to northern Madagascar (Goodman et al. 2009). Within this restricted range, the species is only known to be common in one region, Loky-Manambato, which is characterized by very complex landscape features, including a highly fragmented forest cover (Goodman and Wilme´ 2006). Given the extensive loss of this species’ habitat, there is a need to develop molecular tools to assess the genetic impacts of anthropogenic forest fragmentation on its populations. We present here in the first microsatellite loci isolated for E. carletoni (and for the entire genus and sub-family for that matter), which can be used to achieve this goal (e.g. see De Young and Honeycutt 2005). We followed the established protocol described in Glenn and Schable (2005) to perform microsatellite screening. Total genomic DNA from several frozen tissues was extracted using a DNeasy extraction kit (Qiagen Inc.). Restriction enzyme digests were performed using Rsa I and Xmn I (New England Biolabs) to generate DNA fragments of 300–500 bp. DNA fragments were subsequently ligated with linkers using double stran
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