Nancy A. Schable

Isolating Microsatellite DNA Loci

A series of techniques are presented to construct genomic DNA libraries highly enriched for microsatellite DNA loci. The individual techniques used here derive from several published protocols but have been optimized and tested in our research laboratories as well as in classroom settings at the University of South Carolina and University of Georgia, with students achieving nearly 100% success. Reducing the number of manipulations involved has been a key to success, decreasing both the failure rate and the time necessary to isolate loci of interest. In our lab during the past 3 years alone, these protocols have been successfully used to isolate microsatellite DNA loci from at least 55 species representing three kingdoms. These protocols have made it possible to reduce the time to identify candidate loci for primer development from most eukaryotic species to as little as 1 week.

Five Hundred Microsatellite Loci for Peromyscus

Mice of the genus Peromyscus, including several endangered subspecies, occur throughout North America and have been important models for conservation research. We describe 526 primer pairs that amplify microsatellite DNA loci for Peromyscus maniculatus bairdii, 467 of which also amplify in Peromyscus polionotus subgriseus. For 12 of these loci, we report diversity data from a natural population. These markers will be an important resource for future genomic studies of Peromyscus evolution and mammalian conservation.

Isolation and characterization of microsatellite DNA loci from Ambystoma salamanders

Dean A. Croshaw* Nancy A. Schable Maureen B. Peters & Travis C. Glenn Savannah River Ecology Laboratory, University of Georgia, Drawer E, Aiken, SC 29802, USA; Department of Biological Sciences, University of New Orleans, New Orleans, LA 70148, USA; Department of Biological Sciences, University of South Carolina, Columbia, SC 29208, USA (*Corresponding author: Phone: +803-725-5873; Fax: +803-725-3309; E-mail: croshaw@srel.edu)

Ten microsatellite loci from Northern Bobwhite (Colinus virginianus)

Ecological studies using microsatellite data often require the selection of an optimal marker set for use in parentage and relatedness inference. Commonly, this requires a candidate pool of microsatellite markers from which several are selected to ensure data are acquired efficiently and accurately. We developed 10 microsatellite loci for use with Northern Bobwhite (Colinus virginianus) and tested these loci using individuals collected from two distinct populations in GA and VA. Our new markers yielded seven alleles/locus (range: 2–16) in the Georgia population and six alleles/locus (range: 2–13) in the Virginia population. Exclusionary power of all markers in each population with both parents unknown was >0.98. These microsatellite loci should be combined with previously developed markers to select an optimal set for use in subsequent analyses of parentage and relatedness.

Novel microsatellite markers for the saltmarsh sharp-tailed sparrow, Ammodramus caudacutus (Aves: Passeriformes)

We have developed eight high‐quality microsatellite DNA loci for the saltmarsh sharp‐tailed sparrow and one additional locus with evidence of null alleles. In a sample of 250–350 individuals, the average number of alleles per locus was 14.7 and average observed heterozygosity was 0.80. These loci were tested in three additional species of emberizid sparrows, indicating that more than half of the loci could be useful in other sparrows.

Development and characterization of tetranucleotide microsatellite loci for the American alligator (Alligator mississippiensis)

We isolated and characterized 17 tetranucleotide microsatellite loci in the American alligator, Alligator mississippiensis. Loci were screened across 27 individuals from one population and shown to be polymorphic with the number of alleles per locus ranging from 2 to 12. Polymorphic information content ranged from 0.2 to 0.85, and observed heterozygosity ranged from 0.185 to 0.889. One locus showed significant deviation from Hardy–Weinberg equilibrium, and one pair of loci showed evidence of linkage.