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Journal of Biological Sciences

Year: 2017 | Volume: 17 | Issue: 8 | Page No.: 410-415
DOI: 10.3923/jbs.2017.410.415
Development of Internal PCR Control (IPC) for Human Mitochondrial DNA Typing Kit
Ishar Seri Mirianti , Abdullah Nur Azeelah and Zainuddin Zafarina

Abstract: Background and Objective: Amplification of target sequences has become a necessary technique in molecular research and has been used for various applications such as forensic investigation, genetic tracking, disease diagnosis and much more. The objective of this study was to develop an internal PCR control (IPC) for mitochondrial DNA (mtDNA) typing using allele specific PCR (asPCR) method. Materials and Methods: A total of 5 non-competitive IPC with 300 bp in length were synthesized by incorporating the mtDNA target sequence in the fragment. Both target sequences and IPCs shared similar forward primers. However, reverse primers were designed for each IPC in order to obtain the desired fragment length. Each developed IPC consist of 3 to 6 selected mt DNA SNPs that was used as control to determine the presence of variant in the target sequence, without the need of DNA sequencing. A total of 20 mtDNA SNPs from both coding and control regions were selected for asPCR (16 148, 3552, 16 355, 195, 1872, 1709, 16 108, 16 335, 16 274, 8440, 13 626, 16 291, 9080, 3705, 4491, 146, 1719, 16 093, 3027 and 7684). Results: The IPCs for all selected mtDNA SNPs in asPCR were successfully developed and amplified. The successful amplified IPCs were observed in either wild type lane or variant type lane. Conclusion: Development and incorporation of IPC is important and necessary as these fragments act as control to monitor the performance and amplification results of the asPCR.

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How to cite this article
Ishar Seri Mirianti, Abdullah Nur Azeelah and Zainuddin Zafarina, 2017. Development of Internal PCR Control (IPC) for Human Mitochondrial DNA Typing Kit. Journal of Biological Sciences, 17: 410-415.

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