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APPLICATION OF REAL-TIME PCR TECHNIQUE TO DETERMINE MITOCHONDRIAL COPY NUMBER FROM FFPE SAMPLES
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Abstract
Mitochondria are organelles of eukaryotic cells responsible for energy production. The mitochondrial genome has a circular DNA structure, 16,569 base pairs long, containing 37 genes encoding 13 proteins, 22 tRNAs, and 2 rRNAs. Each mitochondrion can contain hundreds to thousands of copies, depending on the cell type. However, changes in the number of mitochondrial DNA copies (mtDNA-CN) have been shown to be associated with various pathological conditions, including cancer. Formalin-fixed paraffin-embedded (FFPE) tissues are valuable materials for diagnosing and studying cancer pathogenesis. However, DNA extracted from FFPE tissues is heavily degraded due to cross-linking between nucleic acid strands. Real-time PCR is the gold standard method for measuring mtDNA-CN. In this study, we established a simple SYBR Green Real-time PCR method to determine mtDNA-CN from FFPE samples.
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Keywords
Mitochondria, FFPE, Real-time PCR
References
2. Dedhia P, Tarale S, Dhongde G, Khadapkar R, et al. Evaluation of DNA extraction methods and real time PCR optimization on formalin-fixed paraffin-embedded tissues. Asian Pac J Cancer. 2007;8(1):55–59.
3. Christopher B. Jackson, Sabina Gallati, André Schaller. qPCR-based mitochondrial DNA quantification: Influence of template DNA fragmentation on accuracy. Biochemical and Biophysical Research Communications. Volume 423, Issue 3, 2012, 441-447,
4. Bai RK, Chang J, Yeh KT, Lou MA, Lu JF, Tan DJ, et al. Mitochondrial DNA Content Varies with Pathological Characteristics of Breast Cancer. Journal of Oncology. 2011;2011:1–10.
5. Alejandro Alegría Torres J. The mitochondrial DNA copy number used as biomarker. Int J Mol Biol Open Access. 2018;3(3):117–119
6. Birch-Machin MA. Using mitochondrial DNA as a biosensor of early cancer development. Br J Cancer. 2005 Aug;93(3):271–2.
7. Font A, Tort F, Navarro-Sastre A, Cusí V, García-Villoria J, Briones P, Ribes A. Quantitative Analysis of mtDNA Content in Formalin-Fixed Paraffin-Embedded Muscle Tissue. JIMD Rep. 2011;1:125-9.
8. S. Cardoso, Z.S. Quintero-Niño, X. Elcoroaristizabal, I. Guerra-Merino, M.M. de Pancorbo, Mitochondrial DNA analysis of formalin-fixed paraffin-embedded tissue samples: Effect of formalin on DNA stability and its implications in genetic studies. Forensic Science International: Genetics Supplement Series. Volume 3, Issue 1, 2011, e518-e519.