Integration of the AmpFlSTR Identifiler PCR Amplification Kit with SRY-specific primers for gender identification

Dropout of the amelogenin Y-specific allele due to an interstitial deletion of the Yp involving the amelogenin Y locus (AMELY) can cause misidentification of sex genotype with potentially serious consequences in personal identification processes and criminal investigations. Inclusion of additional sex-defining markers in forensic DNA typing kits is therefore advisable. In this study, the co-amplification of the sex-determining region Y (SRY) gene and 16 STR loci included in the AmpFlSTR Identifiler PCR Amplification Kit was evaluated. Combination of SRY and Identifiler primers did not compromise the amplification outcome and generated a 90 bp male-specific SRY fragment, showing a reproducible peak height ratio in comparison with the AMELY peak. The SRY peak was detectable in presence of amounts of template DNA as low as 125 pg, and in mixed samples with a male/female DNA ratio of 1:100.