In this study, the validation of 12 short tandem repeated (STRs) systems is presented in a sample of the population from Antioquia, central region of Colombia. The DNA for the analyses of PCR was obtained of 244 to 546 unrelated individuals. PCR amplification was performed according to the manufacturer's recommendations, using PowerPlex 1.2 System and FFFL Multiplex (Promega). Amplificated fragments were separated by capillary electrophoresis using the ABI Prism 310 Genetic Analyzer (PE Applied Biosystems). Allele frequencies, Hardy-Weinberg equilibrium, observed and expected heterozygosity and genetic disequilibrium were calculated using GENEPOP ver. 3.3 and GDA ver. 1.1 software. Forensic parameters were calculated using the PowerStat ver. 1.2 software (Promega). The 12 of the loci met Hardy-Weinberg expectations and they reached a combined power discrimination higher than 0.999999999999 and a combined power exclusion similar to 0.9999, showing to be a powerful tool for paternity testing, individual identification and forensic applications in Antioquia population.