Development of six-SNPs assay for forensic analysis in European population

Y-chromosomal SNPs analysis show regional specificity useful in forensic investigation for inferring the male genetic background of individuals and population and to predict biogeographical origin of the donor of a crime scene sample. Due to its exclusively paternal inheritance, the Y-chromosome has been extensively used in evolutionary and forensic genetics to investigate the phylogeny and the history of population and their migration. A large scale parsimonious phylogenetic tree representing worldwide Y-chromosome variation has been constructed and comprises major haplogroups. The aim of this study was to set-up six multiplex assays based on SNaPshot kit to identify markers inside major clades of European population. Specifically, we design PCR and minisequencing primers targeting a total of 33 Y-mutations downstream R1*, I*, J2*, G* and E1b1b1* haplogroups. The PCR fragments were chosen to get the shortest product possible in order to improve the performance in degraded samples (amplicons principally ranging from 56 bp to 140 bp). This assay based on a 6-multiplex PCR reaction is a suitable tool for detecting the main European haplogroups in forensic casework and population study.