Methodological Advances in Mitochondrial DNA Analysis for Forensic Genetics

Area of Science:

• Forensic genetics
• Molecular biology
• Human identification

Background:

• Mitochondrial DNA (mtDNA) analysis is vital in forensic genetics for degraded samples.
• Its high copy number and matrilineal inheritance aid in human identification and lineage reconstruction.
• Applications include disaster victim identification, historical cases, and missing persons investigations.

Purpose of the Study:

• To review contemporary methodological approaches for human mitogenome investigation.
• To highlight advancements in DNA extraction, enrichment, and sequencing techniques.
• To assess the impact of these advancements on forensic mtDNA analysis.

Main Methods:

• Review of recent literature on mitochondrial DNA analysis techniques.
• Discussion of advancements in DNA extraction and enrichment.
• Evaluation of Sanger sequencing, Massive Parallel Sequencing (MPS), and Third-Generation Sequencing (TGS) platforms.

Main Results:

• Extraction and enrichment techniques effectively reduce nuclear mitochondrial DNA sequences (NUMTs) and improve fragment recovery.
• Massive Parallel Sequencing (MPS) significantly enhances sensitivity for detecting low-frequency heteroplasmies.
• Third-Generation Sequencing (TGS) enables full-length native molecule sequencing, minimizing PCR amplification biases.

Conclusions:

• Methodological advances are strengthening the reliability of mitochondrial DNA analysis in forensics.
• Ongoing improvements expand the potential of mtDNA analysis for human identification and lineage reconstruction.
• Challenges such as heteroplasmy interpretation and database limitations persist but are being addressed.