Abstract:

Background: The role of mitochondria in cancer, which has a very important role for the cell, especially in energy metabolism and the production of building blocks, is not yet fully understood. The effects of mitochondrial genome variations and haplogroups, that are determined by the combinations of polymorphisms, on different cancers are still investigated. Therefore, we aimed to optimize and use next generation sequencing protocol in cancer research by amplifying the mitochondrial genome. Materials and Methods: Mitochondrial genome was amplified in two separate reactions with Takara LA Taq DNA polymerase from genomic DNAs obtained from glioma tumor samples. A library was constructed using the "Nextera XT DNA Library Prep Kit" and next generation sequencing was performed on MiSeq FGx device. The data were analyzed in terms of mitochondrial variations, heteroplasmy ratios and haplogroups using the "mtDNA Server" database. Results: Next generation sequencing protocol for the mitochondrial genome was optimized. Several variations and their heteroplasmy ratios were detected for the two tumor samples that were analyzed in detail. In addition, haplogroups of these samples were also determined. Conclusions: The impact of mitochondrial variations on cancer biology is still under investigation. Next generation sequencing technologies are a reliable method that can be used in the analysis of tumors especially with low heteroplasmy ratios.

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