Objective of this study was to evaluate microsatellite markers in paternity testing of native cattle breeds in Turkey. Blood samples were collected from Anatolian Black (n=51), Anatolian Grey (n=54), South Anatolian Red (n=51), Native Southern Anatolian Yellow (n=51), East Anatolian Red (n=45) and Zavot (n = 19) cattle. From the blood samples DNA was isolated by using a standard phenol/chloroform method. A total of 20 microsatellite loci were selected from a FAO/ISAG-suggested list. Polymerase chain reaction products were separated by capillary electrophoresis and marker genotypes were determined by fragment analysis. In statistical analyses, allel numbers, observed (Ho) and expected (He) heterozygosities, deviation from Hardy-Weinberg Equilibrium and probability of exclusion (PE) at each microsatellite locus were calculated. A total of 269 different alleles were observed and the mean allele was identified as 13.45. Mean Ho and He values were observed as 0.619-0.852 and 0.669-0.877, respectively. The results indicated that the microsatellite test panel including the most informative 7 loci had total PE value of >0.9999 in each populations and can thereby be used for parentage testing studies of native cattle breeds in Turkey.