Fig. 3

Method and experimental results for the full OCEANS method. a Potential variants in multiple genetic loci of interest are first enriched using multiplex blocker displacement amplification (BDA) [16, 17]. The amplicons are subsequently appended with SAL adapters using PCR and assembled into concatemers. After size selection to remove short concatemers and excess primers, the concatemers are ligated to the Oxford Nanopore adapter bearing a motor protein, and sequenced using the MinION platform. b OCEANS enables confident variant calls of single-base variants at 0.1% VAF without a matched normal sample. The top diagram shows a randomly selected subset of Nanopore Sequencing reads, and the bottom diagram shows the variant read frequency (VRF), the fraction of Nanopore Sequencing reads at each locus that corresponds to the most frequent single-base variant. The forward and reverse primer regions are shaded in gray, and the BDA enrichment region is shaded in green