A Universal Duplex Sequencing Approach for Accurate Detection of Somatic Mutations

Area of Science:

• Genomics
• Molecular Biology
• Biotechnology

Background:

• Accurate detection of rare somatic mutations is crucial for understanding human disease, aging, and environmental impacts.
• Current sequencing methods face limitations in error rates, genome coverage, and DNA input requirements.

Purpose of the Study:

• To develop a novel duplex sequencing protocol (UDSeq) for ultra-accurate detection of rare somatic mutations.
• To enable comprehensive genomic and exomic profiling with minimal DNA input.

Main Methods:

• UDSeq protocol combines random fragmentation, UMI ligation, and quantitative input control.
• Benchmarking performed on human sperm to estimate error rates.
• Application in cell lines and rodent models to profile exposure-specific mutational patterns.

Main Results:

• UDSeq achieves an estimated error rate of ~2.5×10^-9 per base pair.
• Near-complete genome/exome representation is achieved from as little as 100 pg DNA.
• UDSeq captures mutational signatures from heterogeneous populations and reproduces exposure-specific patterns.

Conclusions:

• UDSeq offers a scalable, low-input platform for accurate somatic mutation profiling.
• The protocol provides improved efficiency and cost-effectiveness compared to previous duplex methods.
• UDSeq facilitates cross-species mutational analysis and detailed study of mutational processes.