in the present study, the pedigree and genotype data of 94 individuals were examined for accuracy of parentage allocations and identity test using 12 fluorescent-labelled microsatellite markers in a highly sensitive and accurate ABI system. The panel of 12 markers showed mean PIC value of 0.71 and Shannon index of 1.65 and 7.58 alleles per locus, which suggests that these markers are highly polymorphic and could be useful for parentage control. Based on scoring allele sizes, a total of 91 alleles were observed within the studied population. the highest and lowest number of alleles was observed for tgla227 locus and tgla126 and Bm1818 loci, respectively. the pedigree was considered incorrect in seven (35%) out of all the evaluated progeny, as their genotype did not match their parents. Combined EP value obtained for all loci in both parentage and identification analysis was 0.99, which indicates the high efficiency of the studied marker set and the accuracy of genotyping in ABI systems. Finally, the present findings confirmed the importance of surveying the pedigree structure and efficiency of 12 fluorescent-labelled microsatellite markers in a single multiplex pcr for parentage testing in the sampled holstein cattle population. key words: parentage testing, microsatellite, holstein cattle, pedigree, multiplex pcr, exclusion probability Per capita milk consumption in Iran has been reported to be 73 kg while the average world figure is between 300 and 350 kg. Holstein cattle are the dominant industrial breed for providing main dairy products in Iran.

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