TECHNICAL NOTE Tetranucleotide microsatellite loci from the critically endangered hawksbill turtle (Eretmochelys imbricata) Brian M. Shamblin • Bonnie E. Berry • Denise M. Lennon • Anne B. Meylan • Peter A. Meylan • Mark E. Outerbridge • Campbell J. Nairn Received: 5 July 2012 / Accepted: 7 July 2012 Ó Springer Science+Business Media B.V. 2012 Abstract We describe isolation and characterization of 14 polymorphic tetranucleotide loci from the hawksbill turtle (Eretmochelys imbricata). We identified an average of 14.5 alleles per locus based on screening of 36 individuals captured on foraging grounds in Bocas del Toro Province, Panama, and 9.6 alleles in 13 individuals captured on foraging grounds in Bermuda. Observed heterozygosity ranged from 0.67 to 1.00, with a mean of 0.85 for the Panama foraging aggregation and 0.83 for the Bermuda foraging aggregation. This microsatel- lite suite has a combined non-exclusion probability of identity of 8.26 9 10–23. These markers should be informative in individual and population-focused analyses. Keywords Eretmochelys imbricata Á Hawksbill turtle Á Microsatellite Á Population The hawksbill turtle (Eretmochleys imbricata) is a globally critically endangered marine species. Demand for the ornately patterned carapace scutes, known as tortoiseshell or bekko, by many cultures and over several centuries has sig- nificantly reduced the size and distribution of nesting pop- ulations of hawksbill turtles (Parsons 1972). In the Greater Caribbean region, several major rookeries were extinguished or severely depleted by overharvesting (Meylan 1999; McClenachan et al. 2006). Resolving connectivity among remnant rookeries and between nesting and foraging sites is essential for conservation planning and genetic data from both mitochondrial and nuclear markers are needed. Dinu- cleotide microsatellite loci have been previously isolated from hawksbill turtles (FitzSimmons et al. 1995; Lin et al. 2008; Miro-Herrans et al. 2008). In addition, several tetra- nucleotide microsatellite markers designed from loggerhead turtles amplify well and conform to Hardy–Weinberg equi- librium expectations in hawksbill turtles (Shamblin et al. 2009). However, additional markers are needed for fine- scale studies such as relatedness analyses. We address this need through development of 14 tetranucleotide markers to complement currently existing markers. Genomic DNA was enriched for microsatellite loci using the methods described by Glenn and Schable (2005) with minor modifications. DNA was extracted from blood samples taken from two juvenile green turtles captured while foraging in the Indian River Lagoon east of Sebas- tian, Florida, USA using a Qiagen Ò DNEasy blood and tissue kit. DNA was digested using RsaI endonuclease (New England Biolabs). Double stranded SuperSNX link- ers (Glenn and Schable 2005) were ligated to the digested genomic DNA overnight at 16 °C. Linker-ligated DNA was recovered using PCR on an Applied Biosystems 9700 thermal cycler and verified by agarose gel electrophoresis. Recovered DNA was hybridized to two different mixtures of biotinylated oligonucleotide probes (Integrated DNA Technologies, mix 1: (AAAC) 6 , (AAAG) 6 , (AATC) 6 B. M. Shamblin Á B. E. Berry Á D. M. Lennon Á C. J. Nairn (&) Daniel B. Warnell School of Forestry and Natural Resources, University of Georgia, Athens, GA 30602, USA e-mail: nairn@uga.edu A. B. Meylan Fish and Wildlife Research Institute, Florida Fish and Wildlife Conservation Commission, 100 8th Avenue SE, St. Petersburg, FL 33701, USA P. A. Meylan Natural Sciences, Eckerd College, 4200 54th Avenue S., St. Petersburg, FL 33711, USA M. E. Outerbridge Bermuda Turtle Project, Bermuda Zoological Society, P.O. Box FL 145, FL BX, Flatts, Bermuda 123 Conservation Genet Resour DOI 10.1007/s12686-012-9720-8