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New methods to identify conserved microsatellite loci and develop primer sets of high cross-species utility – as demonstrated for birds

Authors


  • Current addresses: Clemens Küpper, Department of Biology and Biochemistry, University of Bath, Bath BA2 7AY, UK; Ian R. K. Stewart, Department of Biology, 101 Morgan Building, University of Kentucky, Lexington, KY 40506-0225, USA; Kate L. Durrant, School of Biology, University of Nottingham, University Park, Nottingham NG7 2RD, UK; Susannah Bird, Centre for Novel Agricultural Products, Department of Biology, University of York, York YO10 5YW, UK; Ákos Klein, Behaviour Ecology Group, Department of Systematic Zoology and Ecology, Eötvös Loránd, University, Pázmány P. s. 1/c., 1117 Budapest, Hungary; David Martín-Gálvez, Departamento de Ecología Evolutiva y Funcional, Estación Experimental de Zonas Áridas (CSIC), Almería 04001, Spain; Lewis G. Spurgin, School of Biological Sciences, University of East Anglia, Norwich, NR4 7TJ, UK.

Deborah Dawson, Fax: +44 (0)114 222 0002; E-mail: D.A.Dawson@Sheffield.ac.uk

Abstract

We have developed a new approach to create microsatellite primer sets that have high utility across a wide range of species. The success of this method was demonstrated using birds. We selected 35 avian EST microsatellite loci that had a high degree of sequence homology between the zebra finch Taeniopygia guttata and the chicken Gallus gallus and designed primer sets in which the primer bind sites were identical in both species. For 33 conserved primer sets, on average, 100% of loci amplified in each of 17 passerine species and 99% of loci in five non-passerine species. The genotyping of four individuals per species revealed that 24–76% (mean 48%) of loci were polymorphic in the passerines and 18–26% (mean 21%) in the non-passerines. When at least 17 individuals were genotyped per species for four Fringillidae finch species, 71–85% of loci were polymorphic, observed heterozygosity was above 0.50 for most loci and no locus deviated significantly from Hardy–Weinberg proportions.

This new set of microsatellite markers is of higher cross-species utility than any set previously designed. The loci described are suitable for a range of applications that require polymorphic avian markers, including paternity and population studies. They will facilitate comparisons of bird genome organization, including genome mapping and studies of recombination, and allow comparisons of genetic variability between species whilst avoiding ascertainment bias. The costs and time to develop new loci can now be avoided for many applications in numerous species. Furthermore, our method can be readily used to develop microsatellite markers of high utility across other taxa.

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