ONT UL Data
Ultra High Molecular Weight (UHMW) DNA was extracted from approximately 12 L of frozen nucleated blood collected from a Bengalese Finch using the NEB HMW DNA Extraction Kit for Tissues (NEB #T3060), following the ONT recommended gDNA Extraction Protocol (v114_revL_27Nov2022). The protocol was modified by increasing the incubation times for the Proteinase K and RNase A steps to 56 C for 1 hour at 300 rpm. Library preparation was performed using the ONT Ultra-Long DNA Sequencing Kit (SQK-ULK114). The libraries were diluted 3X in EB to account for the high DNA concentration and to facilitate sequencing across multiple flow cells. Two R10.4.1 Flow Cells (FLO-PRO114M) were used for sequencing on the PromethION 48 sequencer (PRO-SEQ048). The flow cells were washed using the ONT wash kit (EXP-WSH004) and reloaded with fresh libraries every 24 hours, for a total runtime of 96 hours. The data was basecalled using dorado0.7.3