Accurate detection of sub-1% frequency somatic mutations by whole-genome sequencing.

Whole-genome sequencing can accurately detect somatic mutations below 1% VAF in high-coverage data, with AVITI and PacBio platforms excelling in different regions.

• Why it matters: Detecting low-frequency somatic mutations is crucial for research and clinical applications, but its reliability across sequencing platforms and genomic regions has not been fully evaluated.
• What they did: The study used primary human fibroblasts, induced pluripotent stem cell lines, and additional iPSC lines from the same donor, employing around 1,360× Illumina and 600× PacBio/AVITI sequencing to benchmark mutation detection.
• The result: Point mutations below 1% VAF and non-homopolymer indels down to 1% VAF were accurately identified, enabling improved detection strategies, though homopolymer indels required VAF >3%, and postmortem samples needed additional filtering.