Isolation and characterization of 18 novel polymorphic microsatellite markers from the Mayan cichlid ( Cichlasoma uropht
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TECHNICAL NOTE
Isolation and characterization of 18 novel polymorphic microsatellite markers from the Mayan cichlid (Cichlasoma urophthalmus) Elizabeth Harrison • Cara N. Love • Kenneth L. Jones • Stacey L. Lance • Joel C. Trexler • Timothy Collins
Received: 26 January 2013 / Accepted: 16 February 2013 / Published online: 27 February 2013 Ó Springer Science+Business Media Dordrecht 2013
Abstract We isolated and characterized 18 microsatellite loci from the Mayan cichlid, Cichlasoma urophthalmus. Loci were screened for 24 specimens from a total of seven sites in south Florida, Mexico, Belize and Honduras. The number of alleles per locus ranged from 3 to 21, observed heterozygosity ranged from 0.208 to 0.875, and the probability of identity values ranged from 0.012 to 0.203. These new loci will provide tools for identifying the source population(s) for the introduction of Mayan cichlids in south Florida and for comparing population genetic structure of Mayan cichlids within and among subpopulations in its native (Central America) and introduced ranges (south Florida). Mayan cichlids are an invasive species in south Florida so identifying source populations may reveal pathways that can be managed to prevent further introductions. Mayan cichlids may also be useful as a model system in which to examine the relationship between introduction history, population genetic diversity, and invasibility. Keywords Cichlasoma Illumina Microsatellite PAL_FINDER PCR primers SSR
E. Harrison (&) J. C. Trexler T. Collins Department of Biological Sciences, Florida International University, 3000 NE 151 St, North Miami, FL 33181, USA e-mail: [email protected] C. N. Love S. L. Lance Savannah River Ecology Laboratory, University of Georgia, Aiken, SC 29802, USA K. L. Jones Department of Biochemistry and Molecular Genetics, University of Colorado School of Medicine, Aurora, CO 80045, USA
The Mayan cichlid is a New World cichlid that is native to the Atlantic slope of Mexico, Belize, Guatemala, Honduras, and Nicaragua (Miller 1966). It was first recorded in the Everglades National Park in 1983 (Loftus and Kushlan 1987); by 1999, it had spread over 200 miles North on both coasts, and is found throughout southern Florida (Adams and Wolfe 2007; Paperno et al. 2008; USGS 2012). Gut content and fish assemblage data indicate that Mayan cichlids negatively impact the densities of small-bodied fishes in estuarine habitats of south Florida including killifish, Sheepshead Minnows and Sailfin Mollies (Loftus 2000; Rehage et al. 2009, Harrison et al. 2013). Total DNA was extracted from one individual of Cichlasoma urophthalmus, using the DNeasy Blood and Tissue kit (QIAGEN) for use in isolation of microsatellite loci. An Illumina paired-end shotgun library was prepared by shearing 1 lg of DNA using a Covaris S220 and following the standard protocol of the Illumina TruSeq DNA Library Kit and using a multiplex identifier adaptor index. Illumina sequencing was conducted on the HiSeq with 100 bp paired-end reads. Five million of the resulti
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