Background: DNA typing in forensic analysis is a useful tool to analyze paternity due to its high discrimination power. Aim: To report the experience of Servicio Medico Legal in Santiago, resolving cases of dubious paternity. Subjects and methods: Four highly polymorphic loci, amplified by polymerase chain reactions, were analyzed in 153 cases of uncertain paternity. The paternity index was calculated for each case. Results: The four genetic markers analyzed provided an exclusion probability of 0.933 for the general population in Santiago. Thirty seven cases were excluded as parents. In 31 cases, the paternity index ranged from 19 to 100, considered as probable paternity and 77 cases had an index of over 100, considered as almost certain paternity. Eight cases had an index between 0.5 and 19, considered as inconclusive. All loci met Hardy-Weinberg expectations and their frequencies were similar to other data from people living in Santiago. Conclusions: The use of these genetic markers proved to be very useful, reliable and with a high exclusion power for paternity analysis