Figure 1. BAC-based longboard nanopore sequencing strategy on the MinION.

(a) Optimized strategy to cut each circular BAC once with transposase results in a linear and complete DNA fragment of the BAC for nanopore sequencing. (b) Yield plot of BAC DNA (RP11-648J18). (c) High-quality BAC consensus sequences were generated by multiple alignment of 60 full-length 1D reads (shown as blue and yellow for both orientations), sampled at random with ten iterations, followed by polishing steps (green) with the entire nanopore long-read data and Illumina data. (d) Circos representation²⁰ of the polished RP11-718M18 BAC consensus sequence. Blue arrowheads indicate the position and orientation of HORs. Purple tiles in yellow background mark the position of the Illumina-validated variants. Additional purple highlight extending from select Illumina-validated variants are used to identify single-nucleotide-sequence variants and mark the site of the DYZ3 repeat structural variants (6 kb) in tandem.