We report autosomal STR data (Identifiler PCR amplification kit) of a total sample of 884 unrelated Mestizos from three different regions of Mexico. The population sample included 309, 313 and 262 individuals from the states of Jalisco (West), Puebla (Center) and Yucatan (Southeast), respectively. Allele distribution and forensic statistical parameters are described. Genotype distribution by locus and two-loci combination was in agreement with Hardy-Weinberg expectations for all 15 STRs. Pairwise comparisons including Mexican populations reported in the literature demonstrated a significant differentiation, principally between North/West with regard to Center/Southeast Mexico. These results increase STR data from previously unreported regions of this country, and constitute a valuable guide in forensic casework for choosing an auxiliary STR database in states where it is not available.