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P233 – Comparative Evaluation of Mitochondrial DNA Extraction Methods from Human Tissues for Forensic Applications
Mitochondrial DNA (mtDNA) has gained increasing attention in forensic science due to its high copy number, maternal inheritance, and heteroplasmy, which provide valuable information for individual identification and kinship analysis, particularly when nuclear DNA is highly degraded or unavailable. However, the efficient recovery of high-quality and intact mtDNA from different types of biological samples remains a critical challenge, limiting its broader application in forensic practice, especially in cases involving degraded or trace biological evidence. This study aimed to systematically evaluate and compare three mtDNA extraction methods, including the QIAamp DNA Investigator Kit, the Saint-Bio Cell Mitochondria DNA Isolation Kit, and an optimized ancient DNA (aDNA) extraction protocol, under standardized experimental conditions. These methods were applied to three types of human tissues, namely peripheral blood, saliva, and hair shafts without follicles, to assess their performance across different sample conditions, including both relatively high-quality and low-template biological materials. The extracted DNA was evaluated using NanoDrop 2000 spectrophotometry and Qubit fluorometric quantification to determine DNA yield and purity. In addition, PCR amplification and agarose gel electrophoresis were conducted to assess DNA integrity and amplification efficiency. The results demonstrated that the optimized aDNA extraction method consistently outperformed the other two methods in terms of DNA yield and amplification success. Notably, this method showed more stable and reproducible performance across different tissue types, indicating its robustness and adaptability under varying sample conditions. In contrast, the commercial extraction kits exhibited relatively lower efficiency, particularly in samples with potentially degraded or low-template DNA. In conclusion, the optimized aDNA extraction protocol provides a more effective and reliable approach for recovering mtDNA from human tissues, especially under challenging forensic conditions. This study offers valuable insights into mtDNA extraction strategies and contributes to the development of a more universal and robust workflow, facilitating downstream applications such as comprehensive forensic mitochondrial genome analysis in forensic investigations.
Authors
- Hong Zhu (Sichaun University, China)
- Dongxu Ma (Sichaun University, China)
- Shuyuan Zhang (Sichaun University, China)
- Jing Liu (Sichaun University, China)
- Zheng Wang (Sichaun University, China)
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