Microsatellite DNA polymorphism and its usefulness for pedigree verification in Simmental cattle from Serbia

Abstract

The polymorphism of 11 microsatellite DNA markers (TGLA227, BM2113, TGLA53, ETH10, SPS115, TGLA126, TGLA122, INRA23, ETH3, ETH225, BM1824) was analyzed in order to assess the usefulness of these markers for pedigree verification in the Serbian population of Simmental cattle. A total of 86 animals were involved in the study. Genomic DNA was extracted from sperm and blood and amplified in a multiplex PCR using primers from the commercial 'StockMarks for Cattle® Bovine Genotyping Kit'. Analyses were done with automated DNA sizing technology. A total of 92 alleles were detected. The mean number of alleles (nA) per locus was 8.364, the mean value of polymorphism information content (PIC) was 0.73, and the mean frequency of the most frequent allele (FNA) was 0.372. The most polymorphic microsatellite loci were: TGLA53 (14 alleles, PIC=0.88, FNA=0.235), TGLA227 (11 alleles, PIC=0.82, FNA=0.274), INRA23 (11 alleles, PIC=0.86, FNA=0.167), BM2113 (9 alleles, PIC=0.84, FNA=0.330). Both combined power of discrimination (CPD) and combined power of exclusion (CPE) for the whole set of studied markers were 0.999. In conclusion, the polymorphism of 11 microsatellite DNA markers observed in this study indicate its usefulness for paternity testing and pedigree verification in Simmental cattle from Serbia.

U ovom radu analiziran je polimorfizam 11 mikrosatelitskih DNK markera (TGLA227, BM2113, TGLA53, ETH10, SPS115, TGLA126, TGLA122, INRA23, ETH3, ETH225, BM1824) u cilju procene upotrebljivosti tih markera u verifikaciji pedigrea Simentalskih goveda u Srbiji. Analizirano je ukupno 86 grla. Genomska DNK je izolovana iz sperme i krvi. Za amplifikaciju je primenjen multiplex PCR korišćenjem prajmera komercijalnog seta 'StockMarks for Cattle® Bovine Genotyping Kit'. Pri analizi rezultata korišćena je tehnologija automatizovanog određivanja veličine DNA fragmenata. Ukupan broj detektovanih alela bio je 92. Srednja vrednost broja alela (nA) po lokusu iznosila je 8,364, srednja vrednost sadržaja polimorfne informacije (PIC) bila je 0,73, dok je učestalosti najčešćeg alela (FNA) u proseku iznosila 0,372. Najveća polimorfnost uočena je kod lokusa TGLA53 (14 alela, PIC=0,88, FNA=0,235), TGLA227 (11 alela, PIC=0,82, FNA=0,274), INRA23 (11 alela, PIC=0,86, FNA=0,167), BM2113 (9 alela, PIC=0,84, FNA= 0,330). Moć diskriminacije i moć isključenja celog seta proučavanih markera iznosila je 0,999. Polimorfizam uočen kod 11 mikrosatelitskih DNK markera u ovom radu ukazuje na mogućnost njihove primene u testiranju očinstva i verifikaciji pedigrea Simentalskih goveda u Srbiji.