The aim of the studies was to evaluate the polymorphism of 24 microsatellite DNA sequences in 4 Polish cattle breeds, i.e. the Whitebacks (BG) –100 animals, the Polish Red (RP) – 60 animals, the Polish Black-and-White (BW) – 50 animals and the Polish Holstein-Friesian of Black-and-White variety (PHF-HO) – 50 animals. The most polymorphic loci of all races were: TGLA53, TGLA227, TGLA122, INRA037 and HEL9, in which 17, 15, 14, 13 and 12 different alleles were identified, respectively. Allele 186-bp at ILSTS005 locus was the most frequent in BG (0.675), allele 101 bp in locus INRA035 was the most frequent in RP and BW (with the value of 0.742 and 0.730, respectively) and 163 bp allele in HEL5 locus of race PHF- -HO (0.637). Using a total of 24 loci, the probability to exclude a wrong parent for each of the races was 0.999.