PZR metoduyla atlarda birey ve ebeveyn tayini

Yıl 2012, Cilt: 28 Sayı: 2, 77 - 81, 01.06.2012

Mehmet Nizamlıoğlu Ercan Kurar Zafer Bulut Şeref İnal Ferudun Erzurum

Öz

Amaç: Atlarda ebeveyn tayininde kullanılabilecek bir mikrosatellit panelini geliştirmek ve test etmektir.Gereç ve Yöntem: Araştırmada 4 farklı at ırkından 189 adet kan örneği toplandı. Beş adet mikrosatellit markörü kullanılarak, izolasyonu yapılan genomik DNA örnekleri Polimeraz Zincir Reaksiyonu (PZR) metoduyla çoğaltıldı ve PZR ürünleri poliakrilamit jellerde ayrıştırıldı. Standart bir DNA ladder yardımıyla alleller tanımlandı.Bulgular: Lokus başına 9-13 arasında değişen, toplam 53 farklı allel tanımlandı. Gözlenen (Ho) ve beklenen heterozigotluk (He) değerleri sırasıyla 0.496-0.880 ve 0.800-0.851 arasında bulundu. Polymorphism Information Content değerleri ise 0.774-0.832 arasında gözlemlendi. Dışlama gücü (DG) değerleri 0.619-0.702 arasında gözlemlenirken, toplam DG 0.99060 olarak bulundu. Öneri: Kullanılan test panelinin Türkiye’de ebeveyn tayininde kullanılabileceği ve hatalı sonuçları azaltmak için kullanılan mikrosatellit sayısının arttırılmasının faydalı olacağı kanaatine varıldı.

Anahtar Kelimeler

At, mikrosatellit, ebeveyn testi

Individual and parentage testing in horses by PCR methodology

Öz

Aim: The objective of this study was to develop and test a microsatellite panel for parentage analysis in horses. Materials and Methods: A total of 189 blood samples were collected from four different horse breeds in Turkey. We selected five horse microsatellite loci and used to amplify genomic DNA by polymerase chain reaction (PCR). The resulting PCR products were separated on polyacrylamide gels. Allele identification was conducted based on their base-pair size by comparing a size standard. Results: A total of 53 alleles was determined ranging from 9 to 13 at each locus. The observed heterozygosity (Ho) and expected heterozygosity (He) were ranged from 0.496 to 0.880 and from 0.800 to 0.851, respectively. Polymorphism information content (PIC) values were observed between 0.774 and 0.832. Power of exclusion (PE) at each microsatellite locus ranged from 0.619 to 0.702, resulting in a total PE value of 0.99060. Conclusion: These results indicate that this set of microsatellite is useful for horse parentage testing in Turkey. Due to possible high level of inbreeding in some breeds, the use of increased number microsatellite loci will thereby be appropriate for avoiding a false parenting and misidentification.

Anahtar Kelimeler

Horse, microsatellite, parentage

Kaynakça

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