SREL Reprint #2427

Dinucleotide microsatellite loci in a migratory wood warbler (Parulidae:  Limnothlypis swainsonii) and amplification among other songbirds

KEVIN WINKER¹, TRAVIS C. GLENN², and GARY R. GRAVES³

¹University of Alaska Museum, 907 Yukon Drive, Fairbanks, Alaska 99775, USA
²Savannah River Ecology Laboratory, Drawer E, Aiken, SC 29802; & Department of Biological Science, University of South Carolina, Columbia, SC 29208, USA
³National Museum of Natural history, Laboratory of Molecular Systematics, MSC MRC 534, Smithsonian Institution, Washington, D.C. 20560

Development of conservation and management plans can be aided by genetic data, and the utility of microsatellites for studies of genetic variability, population structure, gene flow, and relatedness makes these loci appropriate for studies of declining natural populations. Wood warblers (Parulidae), a New World family of 115 species of 25 genera, have played an important role in studies of avian ecology, evolution, hybridization, and behaviour. Also, 7% of the species are threatened or endangered (Collar et al. 1992), and populations of other species are locally threatened or declining.

We developed polymerase chain reaction (PCR) assays of microsatellite loci for Swainson's Warbler (Limnothlypis swainsonii), the only species in the genus Limnothlypis and among the rarest wood warblers in North America (Brown & Dickson 1994). Our PCR primers were tested for their ability to amplify orthologous loci in 19 other wood warblers and five other species of three different families.

Protocols used to obtain and score microsatellites (Glenn 1997) involved ligating 300-700 bp fragments of genomic DNA into a plasmid and enriching for dinucleotide microsatellites using d(AG)₁₂ and d(AC)₁₂ or d(AG)₁₂ and d(TG)₁₅ (Ostrander et al. 1992). Transformed bacterial colonies were screened by hybridization with radioactively labelled d(AG)₁₂ and d(TG)₁₅. Sixty positive clones were sequenced following Meeker et al. (1993) or Rouer (1994) using Sequenase 2.0 (US Biochemical). PCR primers were designed from sequences flanking the repetitive elements. Genomic DNA was extracted using a diatomaceous earth/guanidine thiocyanate extraction protocol (Carter & Milton 1993) and diluted to 20 ng/µL for use in PCR experiments.

SREL Reprint #2427

Winker, K., T.C. Glenn, and G.R. Graves. 1999. Dinucleotide microsatellite loci in a migratory wood warbler (Parulidae:  Limnothlypis swainsonii) and amplification among other songbirds. Molecular Ecology 8:1553-1556.

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