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.