Human sample from Homo sapiens
Ultra-high molecular weight (uHMW) DNA was prepared from either 1.6 mL of freshly collected blood or 6 million cultured cells DNA as described in the ONT Ultra-long DNA sequencing kit procedure version November 28th, 2022 with a Monarch HMW DNA Extraction Kit for Tissue T3060. Ultra-Long DNA libraries were prepared using the ONT Ultra-Long DNA Sequencing Kit (SQK-ULK114). Up to 40 g of UL DNA were fragmented, had rapid adaptors added, and cleaned by precipitation according to the ONT protocol and eluted in 300 L of elution buffer overnight. One PromethION R10.4.1 flow cell was used to sequence each sample.  90 L of the QCd UL library was loaded onto a single PromethION R10.4.1 flow cell following ONT procedure with reagents from SQK-ULK114. Adaptive sampling sequencing runs were set for 72 hours with a minimum read length of 1000 bp and FAST live basecalling. Adaptive sampling was enabled and a reference file containing MHC and KIR haplotype sequences was submitted to the control software. Libraries were reloaded every 24 hours until a full run time of 72. After sequencing was completed, ONT reads were rebasecalled using Dorado (v.0.5.3) and the super accurate (SUP) model (v4.3). Fastq sequences were generated for reads with a minimum quality score of Q10.