The genetic characterization of 117 peach and nectarine cultivars (Prunus persica (L.) Batsch) using microsatellite (SSR) markers is presented. Analyzed genotypes include the complete list of cultivars under intellectual property (IP) protection in Chile. One hundred and two out of the 117 cultivars under study could be identified using only 7 SSRs. Other 5 cultivars were differentiated using 3 additional markers, but 5 pairs of genotypes were not differentiable. The average expected heterozygosity for the set of markers was 0.55, ranging from 0.28 in BPPCT-008 to 0.81 in CPPCT-022, with an F value of 0.37. A Neighbor-Joining dendrogram showed that, with few exceptions, peaches and nectarines clustered separately. These results are the basis for the development of a fingerprinting protocol for the unequivocal identification of most of the peach and nectarine cultivars officially registered in Chile.