Molecular Gender Determination of Ancient Human from Malay Peninsular

Problem statement: DNA samples from fourteen modern human bloods (seven males and seven females) and two ancient skeletal samples excavated from Kalumpang Island and Dungun, Peninsular Malaysia were subjected for molecular genders determination using specific primers of human AMELX and AMELY. Approach: A standard multiple PCR mixture with forward primer and either a human X-specific reverse primer for AMELX or a human Y-specific reverse primer for AMELY amplifications were used to assess the presence of these genes in male and female samples. Results: PCR amplification of a modern male sample yielded 329 and 235 bp bands, whereas a modern female sample only 329 bp band. The Kalumpang Island sample produced two positive bands of AMELY and AMELX. After reamplification of the Dungun sample, only an AMELY band was visible. All amplified bands were cloned into TA plasmid and sequenced. BLAST analysis showed that the 329 bp band is AMELX, while the 235 bp product is AMELY. Conclusion/Recommendations: The skeletal remains of both Kalumpang Island and Dungun samples from west and east of Peninsular Malaysia respectively, are males.