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

Molecular Ecology Resources - Tập 10 Số 3 - Trang 475-494 - 2010
Deborah A. Dawson1, Gavin J. Horsburgh1, Clemens Küpper1, Ian Stewart1, Alexander D. Ball1, Kate L. Durrant1, Bengt Hansson2, Ida Elizabeth Bacon3, Susannah Bird1, Ákos Klein1, Andrew P. Krupa1, Jinwon Lee1, David Martín‐Gálvez1, Michelle Simeoni1, Gemma Smith1, Lewis G. Spurgin1, Terry Burke1
1Department of Animal and Plant Sciences, University of Sheffield, Sheffield S10 2TN, UK
2Department of Animal Ecology, Ecology Building, Lund University SE-223 62 Lund, Sweden
3Institute of Evolutionary Biology, School of Biological Sciences, King's Building, University of Edinburgh, Edinburgh, EH9 3JT, Scotland, UK

Tóm tắt

AbstractWe 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 finchTaeniopygia guttataand the chickenGallus gallusand 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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Molecular Ecology Resources

Tập 10 Số 3

475-494

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