The duplicated Y-STR DYS385 is one of the most informative markers on the Y-chromosome. It consists of two copies-DYS385a and DYS385b-that are located close to the inner borders of palindrome P4. The application of standard PCR protocols for DYS385 results in the simultaneous amplification of both copies. Hence, an explicit assignment of the alleles to their loci is not possible which causes a loss of information. Recently, a PCR strategy was published that allows the separate analysis of DYS385a and b, which we followed with modifications in our study on 133 unrelated Caucasian men from Tirol (Austria). Additionally all markers of the minimal haplotype (including DYS385) were amplified using standard PCR protocols. The non-allele-discriminating amplification strategy for DYS385 allowed the discrimination of 27 different genotypes (GD=0.8616). By the separate analysis of the two DYS385 copies, this number increased to 34 (GD=0.9185). However, the cumulative haplotype diversity of the complete minimal haplotype was 0.9951, regardless of the amplification strategy for DYS385. This indicates that in the context of the minimal haplotype, the separate analysis of DYS385a and b has no or only a marginally positive effect on the differentiation of paternal lineages.