The allele and genotype frequency values of the three tetranucleotide short tandem repeat (STR) loci, D7S820, D13S317 and D16S359, were analysed in blood samples of 25 unrelated randomly selected individuals in the National Capital District, Papua New Guinea. Gene-Print Silver-STR III Multiplex kit (Promega Corp., Medison, WI, USA) was used for the PCR amplification in GeneAmp®PCR System 9700 thermal cycler (Applied iosystems). Data analysis was carried out using the PowerStatsV12.xl workbook template obtained from Promega Corporation. The three STR loci were in Hardy-Weinberg equilibrium. Five alleles (9 – 13) were identified for D16S539, five alleles (8 – 12) for D7S820 and six alleles (8, 9, 11 – 14) for D13S317. No new or microvariant alleles were observed. The most frequent genotypes for D16S539 were 11-11 and 11-12; for D7S820 were 10-11 and 12-12; for D13S317 was 8-12. Observed Heterozygosity was highest in D13S317 (0.880). The combined power of discrimination was 0.99733 and the combined power of exclusion was 0.9363. The data suggests that the three loci are useful for identity testing, forensics and for solving paternity cases among the population in National Capital District, Papua New Guinea.