In this work we present results on DYS434, DYS437, DYS438 and DYS439 loci studied in three population samples from North Portugal (N: 69), Macao (N: 59) and Mozambique (N: 64). Gene and haplotype diversity values are presented and compared. Gene diversity values ranged from 0.0290 to 0.0620 for DYS434; 0.0615 to 0.5959 for DYS437; 0.4906 to 0.6424 for DYS438; 0.5873 to 0.7038 for DYS439. The highest average gene diversity was found in Macao and the lowest in Mozambique. Haplotype diversity in North Portugal, Macao and Mozambique was calculated to be 0.925, 0.928 and 0.787, respectively. Allele and haplotype frequency distributions for all markers were compared between the three populations. At the haplotype level all the differences between pairs of samples were significant. Sequencing analysis was performed for all alleles found in the four loci. Identical flanking sequences were observed with most of the alleles differing only by the repeat number. However, two variations were found in allele 10 of DYS438: one in a Macao sample presenting a TTTTA before the last TTTTC repetition and one in a Portuguese sample showing a base substitution (A/C) at position 7 downstream from the tandem repeat. A base substitution (C/T) at position 3 upstream from the repeat motif was also found in allele 14 of DYS437 in a sample from Mozambique.