Abstract
DNA from non-invasively collected samples may be used to identify individuals for monitoring birds. Here we describe 36 novel microsatellite loci from Bean Goose (Anser fabalis) identified using a next-generation sequencing approach. From 34 variable loci, we selected 12 tetranucleotide loci with short amplicon sizes, combined these into two multiplex PCR sets, and included a sex-typing marker. In 31 Bean Geese from a population in central Norway, we found 4–10 alleles per locus and observed heterozygosity ranging from 0.58 to 0.87. The combined probability of identity for the 12 loci was 6.5 × 10−13. These novel microsatellite loci and their multiplex PCR assays will be useful for genetic analyses of the Bean Goose, including typing of non-invasive samples such as molted feathers. Cross-species application of the two multiplex PCR assays revealed that all 12 loci amplified successfully in four other Anser species.
Zusammenfassung
Isolierung, Charakterisierung und Entwicklung einer Multiplex-PCR Methode für Mikrosatellitenloci von Saatgänsen ( Anser fabalis )
Für die Identifizierung von Individuen kann DNA aus nicht-invasiv gesammelten Proben zum Monitoring von Vögeln genutzt werden. In dieser Studie beschreiben wir 36 neue Mikrosatelliten-Genorte von Saatgänsen (Anser fabalis), die im Rahmen einer Next-Generation Sequenzierungsmethode identifiziert wurden. Aus 34 variablen Stellen selektierten wir 12 Tetranucleotidloci mit kurzen Amplikonlängen, kombinierten diese in zwei Multiplex-PCR Sets und fügten einen Geschlechtsbestimmungsmarker hinzu. Bei 31 Saatgänsen aus einer Population in Zentralnorwegen fanden wir 4–10 Allele pro Ort und stellten Heterozygositäten von 0.58–0.87 fest. Die kombinierte Wahrscheinlichkeit der Identität für die 12 Orte war 6.5 × 10−13. Diese neuen Mikrosatellitenorte und ihre Multiplex-PCR Methoden einschließlich der nicht-invasiven Probennahme wie beispielsweise Mauserfedern, können nützlich für genetische Analysen bei Saatgänsen sein. Die Anwendung der zwei Multiplex-PCR Methoden auf andere Arten zeigten, dass sich alle 12 Genorte erfolgreich bei vier anderen Anser-Arten amplifizierten.
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Acknowledgments
We are grateful to Svein Karlsen and Øyvind Spjøtvoll for providing A. f. fabalis shed feathers, and the Natural History Museum in Oslo for providing samples from A. albifrons (NHMO-BI-26959/1-B and NHMO-BI-35338/1-B), A. erythropus (NHMO-BI-18687/1-B, NHMO-BI-18688/1-B and NHMO-BI-18689/1-B) and A. f. fabalis (NHMO-BI-26976/1-B and NHMO-BI-26977/1-B). We thank Deborah A. Dawson for kindly providing the sex-typing markers. Financial support was received from the County administration in Nord-Trøndelag and the Norwegian Environment Agency. Permissions for Bean Goose catching and tissue sampling were approved by the Norwegian Animal Research Authority (NARA, ID 1775 and ref. 2011/64833) and the Norwegian Environment Agency (ref. 2012/1756).
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Kleven, O., Kroglund, R.T. & Østnes, J.E. Isolation, characterization and multiplex PCR development of Bean Goose (Anser fabalis) microsatellite loci. J Ornithol 157, 641–646 (2016). https://doi.org/10.1007/s10336-015-1309-z
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DOI: https://doi.org/10.1007/s10336-015-1309-z