Biotechnology1682-296x1682-2978Asian Network for Scientific Information10.3923/biotech.2018.151.157Mirianti IsharSeri Rus DinRus Dina 32018173Background and Objective: Human DNA is categorized into nuclear DNA (nDNA) and mitochondrial DNA (mtDNA). Mutation occurred in these two types of human DNA can be utilized for the purpose of medical reports, molecular genetic analysis, haplogroup variations, forensic crime search and many others. The polymorphisms were selected from mtDNA in both coding and control regions. Main objective of this project was to produce a simple, cost effective yet robust typing method for human identification using allele specific PCR (asPCR) specifically designed for southeast Asian population. Materials and Methods: A total of 60 subjects with ethical approval were randomly collected with 30, 20 and 10 of them represented Malay, Chinese and Indian population. Polymerase Chain Reaction (PCR) was carried out using 25 sets of primers, that amplify the fragments bearing the selected SNPs. In the second round PCR, two types of allele specific primers labelled as wild type allele specific primer (wtASP) and variant type ASP (vtASP) were used to amplify both Cambridge Reference Sequence (CRS) and variant sequence. The PCR was used for first round and for the second round, the as PCR was applied. Finally, the amplified products were directly viewed using UV light. Results: Overall, out of 30 selected SNPs, the designed ASP managed to amplify only 20 SNPs. The selected SNPs in this study were SNP 146, 195, 1709,1719, 1872, 3705, 3027, 3552, 4491, 7684, 9080, 8440, 13626, 16108, 16291, 16274, 16355, 16093, 16335 and 16148 that reported to belong to macrohaplo group M, B, F, E and N. All the amplified products of selected mtSNPs were observed in wild type lane except for SNP 195 that the amplified product was observed in variant lane. Conclusion: The variant allele managed to be amplified with simple technique yet robust to be brought on site. Generally, mutation found using this technique may narrow down the individual and also population hence it is beneficial in cases such as forensic crime and mass disaster.]]>Asari, M., S. Watanabe, K. Matsubara, H. Shiono and K. Shimizu,20093868590Inagaki, S., Y. Yamamoto, Y. Doi, T. Takata and T. Ishikawa et al.,20041444557Boonyarit, H., S. Mahasirimongkol, N. Chavalvechakul, M. Aoki and H. Amitani et al.,201411166173Hoong, L.L. and K.C. Lek,2005137985Senafi, S., S.H.Z. Ariffin, R.D.R. Din, R.M. Abdul Wahab, I.Z.Z. Abidin and Z.Z. Ariffin,201414197200Deng, L., B.P. Hoh, D. Lu, R. Fu and M.E. Phipps et al.,201413311691185Ricaut, F.X., H. Razafindrazaka, M.P. Cox, J.M. Dugoujon and E. Guitard et al.,20092009Sun, C., Q.P. Kong, M.G. Palanichamy, S. Agrawal and H.J. Bandelt et al.,200623683690Maji, S., S. Krithika and T.S. Vasulu,200988127139Soares, P., F. Alshamali, J.B. Pereira, V. Fernandes and N.M. Silva et al.,20113 within and out of Africa.]]>29915927Hill, C., P. Soares, M. Mormina, V. Macaulay and W. Meehan et al.,20062324802491Palanichamy, M.G., C. Sun, S. Agrawal, H.J. Bandelt and Q.P. Kong et al.,200475966978Li, H., X. Cai, E.R. Winograd-Cort, B. Wen and X. Cheng et al.,2007134481488Kong, Q.P., H.J. Bandelt, C. Sun, Y.G. Yao and A. Salas et al.,20061520762086Mirianti, I.S., Abdullah Nur Azeelah and Z. Zafarina,201717410415Din, R.D.R., S.H.Z. Ariffin, S. Senafi, R.M.A. Wahab and I.Z.Z. Abidin,20144315231535Ariffin, S.H.Z., R.M.A. Wahab, Z. Zamrod, S. Sahar, M.F.A. Razak, E.J. Ariffin and S. Senafi,2007152731Pan, W.L., J.L. Hu, Y. Fang, Q. Luo and G. Xu et al.,2013194277279Wangkumhang, P., K. Chaichoompu, C. Ngamphiw, U. Ruangrit, J. Chanprasert, A. Assawamakin and S. Tongsima,20072007Vallone, P.M., R.S. Just, M.D. Coble, J.M. Butler and T.J. Parsons,2004118147157Clermont, O., J.K. Christenson, A.S. Daubie, D.M. Gordon and E. Denamur,2014Escherichia coli B2 sub-typing, a rapid and easy to perform substitute of multilocus sequence typing.]]>1012427Scheen, A.C., B.E. Pfeil, A. Petri, N. Heidari, S. Nylinder and B. Oxelman,201212128135Cook, L., K. Diem, W. Kim, J.D. Scott and K.R. Jerome,20125041444146Wang, H., J. Jiang, B. Mostert, A. Sieuwerts and J.W. Martens et al.,2013156269Kline, M.C., P.M. Vallone, J.W. Redman, D.L. Duewer, C.D. Calloway and J.M. Butler,200550377385Brandstatter, A., T.J. Parsons and W. Parson,20041261422424Pionzio, A.M. and B.R. McCord,201495560Abdulmawjood, A., S. Roth and M. Bulte,2002Escherichia coli O157 by polymerase chain reaction.]]>16335339