Non size-selected genomic DNA library from single individual
DNA isolated from single female using modified Qiagen MagAttract HMW kit. 100 ng unsheared DNA was used to construct SMRTbell library using Express Prep kit v2.0 (Pacific Biosciences). DNA damage repair, end polishing and tailing with A-overhang performed. Following ligation with T-overhang SMRTbell adapter, library was purified with two AMPure PB beads steps to remove small fragments. Sequencing primer v4 and Sequel DNA Polymerase 3.0 were annealed and bound, respectively. Library diffusion loaded at 5-6 pM on-plate concentration. Sequencing performed with Sequel Sequencing Kit 3.0, 1200-minute movies with 120-minute pre-extension and Software v6.0.