Detection of mitochondrial DNA polymorphisms from human hair shafts and formalin fixed tissue using whole genome amplification

Objective: The objective of this study was to detect mitochondrial DNA polymorphisms from human hair shafts and formalin-fixed tissues using whole genome amplification. Design: A study of forensic medicine. Material and Methods: All of the hairs were taken from the human heads of 9 collaborators (8 healthy males in their 10-40s and 1 healthy female in her 20s), from whom informed consent for the study was obtained. DNA was extracted using QIAamp DNA mini kit. Human hearts, livers, or spleens excised by autopsy and preserved with 10% formalin solution for 7-17 years were used. After genome extension by primer extension preamplification (PEP)-PCR, the mitochondrial DNA (mtDNA) of samples was analyzed in two areas of HVR1, 2. Results: The applicability of PEP-PCR to mitochondrial DNA polymorphism analysis (HVR1, 2) in hair shafts and hearts, livers, and spleens preserved in formalin fixative for a maximum of 17 years was investigated. From hair shafts, HVR1, 2 of the mtDNA areas was amplified in the whole genome amplification product of PEP-PCR. The sequences were successfully analyzed, and were identical to the positive control DNA. Conclusion: whole genome amplification is effective method for analysis of mitochondrial DNA polymorphism from hair shafts and formalin fixed tissues.