This protocol describes a strategy for analyzing phylogenetic Y-SNPs in a hierarchical multiplex assay by utilizing the SNaPshot� Multiplex System. Step by step, the protocol assists in the appropriate selection of SNPs, the primer design, the set up of PCR/SBE reactions as well as in the analysis of the results. Furthermore, a forensic approach is highlighted, in which the most probable ancestry of an unknown male DNA is inferred by the geographical distribution of the assigned Y-SNP haplogroup.