Error-corrected Duplex Sequencing of human TK6 cells exposed to N-ethyl-N-nitrosourea

Error-corrected Duplex Sequencing (DuplexSeq) can correct errors introduced during DNA sample handling and PCR amplification through generating consensus sequences by labeling each strand of a DNA duplex with a unique molecular identifier. This error correction can decrease the error rate of conventional NGS by approximately 10,000x. The DuplexSeq human mutagenicity panel contains 20 2.4-kb target regions across 20 autosomal chromosomes. The panel is representative of the whole genome in terms of % GC-content and relative proportions of coding and non-coding regions and can be used to measure mutation frequencies (MF) in biological samples or cultured cells. In this study, we investigated the utility of DuplexSeq to quantify induced MF and spectrum in human lymphoblastoid TK6 cells exposed to a prototypical DNA alkylating agent, N-ethyl-N-nitrosourea (ENU). In addition, we explored appropriate experimental parameters for this application, and assessed inter-laboratory reproducibility.