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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
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