Pentanucleotide tandem repeat markers are of interest for forensic science because they may present less stutter in electrophoretic patterns. We focused on the analysis of the DNA sequence for each allele at two pentanucleotide STR loci, D6S957 and D10S2325, in order to understand their structures in the human genome and to construct allelic ladders, necessary for forensic DNA typing. In order to evaluate the forensic applicability of both pentanucleotide tandem repeat loci and to construct a preliminary database, the genotype distributions and allele frequencies in different ethnic groups were investigated. The population samples included Caucasians (Germans) and Asians (Chinese). The Amp-FLP technique was employed for DNA typing. An example of each allele and new alleles were sequenced. Allele determination for each pentanucleotide STR locus was carried out by comparison with a sequenced allelic ladder made in-house. Both pentanucleotide STR markers provided easily interpretable results. No evidence of deviation from Hardy-Weinberg equilibrium was observed. In 64 confirmed father/mother/child trios, no mutation event was observed. Using a maximum likelihood method, the mutation rate at both STR loci was indirectly estimated as 2.5 10