Oxford Nanopore Duplex sequencing of HG002
ONT Duplex  sequencing: High molecular weight DNA was extracted from cells using NEB Monarch HMW DNA Extraction Kit for Tissue (NEB T3060). Isolated DNA was then sheared using the Diagenode Megaruptor 3, DNAFluid+ Kit (E07020001). The size of sheared DNA fragments were analyzed on an Agilent Femto Pulse System using the Genomic DNA 165kb Kit (FP-1002-0275). The fragment size distribution of post-sheared DNA had a peak at approximately 50kbp. Small DNA fragments were removed from the sample using the PacBio SRE kit (SKU 102-208-300). Library preparation was carried out using Oxford Nanopore Technologies Ligation Sequencing Kit V14 (SQK-LSK114). PromethION high duplex flow cells were used for sequencing on PromethION 48 sequencer. Three libraries were prepared per flow cell. Flow cells were washed using ONTs Flow Cell Wash Kit (EXP-WSH004) and reloaded with fresh library every 24 hours for a total sequencing runtime of 72 hours.