Development of internal PCR control (IPC) for human mitochondrial DNA typing kit

Seri Mirianti Ishar, Abdullah Nur Azeelah, Zainuddin Zafarina

Research output: Contribution to journalArticle

1 Citation (Scopus)

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.

Original languageEnglish
Pages (from-to)410-415
Number of pages6
JournalJournal of Biological Sciences
Volume17
Issue number8
DOIs
Publication statusPublished - 1 Jan 2017

Fingerprint

DNA Fingerprinting
Mitochondrial DNA
Polymerase Chain Reaction
Alleles
Single Nucleotide Polymorphism
Forensic Genetics
Inborn Genetic Diseases
DNA Sequence Analysis
DNA

Keywords

  • Allele specific PCR
  • Internal PCR control
  • Mitochondrial DNA SNPs
  • mtDNA typing
  • Synthetic DNA

ASJC Scopus subject areas

  • Molecular Medicine
  • Cell Biology

Cite this

Development of internal PCR control (IPC) for human mitochondrial DNA typing kit. / Ishar, Seri Mirianti; Azeelah, Abdullah Nur; Zafarina, Zainuddin.

In: Journal of Biological Sciences, Vol. 17, No. 8, 01.01.2017, p. 410-415.

Research output: Contribution to journalArticle

Ishar, Seri Mirianti ; Azeelah, Abdullah Nur ; Zafarina, Zainuddin. / Development of internal PCR control (IPC) for human mitochondrial DNA typing kit. In: Journal of Biological Sciences. 2017 ; Vol. 17, No. 8. pp. 410-415.
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