A full-length mtDNA dataset for studying genetic variations across generations and complex family structures

Area of Science:

• Genomics
• Molecular Biology
• Bioinformatics

Background:

• Mitochondrial DNA (mtDNA) analysis is vital for disease research, evolution, and lineage tracing.
• Current sequencing methods struggle with nuclear mitochondrial sequences (NUMTs), causing ambiguous results.
• Existing techniques amplify short mtDNA fragments, failing to fully address NUMT interference.

Purpose of the Study:

• To develop a novel third-generation sequencing (TGS) method for full-length mtDNA amplification.
• To overcome NUMTs artifacts in mtDNA sequencing using a single primer pair approach.
• To enable accurate and comprehensive mtDNA characterization for various research fields.

Main Methods:

• Utilized a nanopore-based TGS approach with a single primer pair for full-length mtDNA amplification.
• Employed the QITAN TECH QNome-3841hex platform for sequencing 106 samples from eight family pedigrees.
• Achieved complete mtDNA coverage and high mapping rates for robust genome characterization.

Main Results:

• The method successfully circumvented NUMTs artifacts, providing unambiguous mtDNA sequence data.
• Achieved 100% genome coverage with an average mapping rate of 99.96% across all samples.
• Generated a comprehensive dataset suitable for diverse genetic analyses, including familial structures.

Conclusions:

• The developed nanopore TGS method offers a superior alternative for mtDNA sequencing, effectively mitigating NUMT interference.
• This technique enhances the accuracy of mtDNA mutation detection, mitochondrial genetics, and population studies.
• The findings support advancements in mtDNA sequencing technologies, ancestry tracing, and forensic identification.