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Significance The detection of rare mutations in clinical samples is essential to the screening, diagnosis, and treatment of cancer. Although next-generation sequencing has greatly enhanced the sensitivity of detecting mutations, the relatively high error rate of these platforms limits their overall clinical utility. The elimination of sequencing artifacts could facilitate the detection of early-stage cancers and provide improved treatment recommendations tailored to the genetic profile of a tumor. Here, we report the development of BiSeqS, a bisulfite conversion-based sequencing approach that allows for the strand-specific detection and quantification of rare mutations. We demonstrate that BiSeqS eliminates nearly all sequencing artifacts in three common types of mutations and thereby considerably increases the signal-to-noise ratio for diagnostic analyses.

Bisulfite-converted duplexes for the strand-specific detection and quantification of rare mutations