A duplex sequencing approach for high-sensitivity detection of genome-edited plants.

In this paper, we have evaluated a targeted high-throughput massive parallel sequencing approach for detecting single nucleotide mutations or small genomic changes generated by new genomic techniques (NGT). We used unique molecular identifiers (UMIs) for the quantification of the mutant alleles and duplex sequencing to confirm a mutation on both strands to avoid polymerase chain reaction (PCR) artefacts or sequencing miss-calls. We tested the approach in blinded analyses on a set of mixed NGT-modified tomato lines and identified each single nucleotide mutation or small insert/deletion (InDel) down to a 0.