PromethION sequencing
Oxford Nanopore Technology sequencing for genome assembly
9 µg of high molecular weight DNA was size selected using the Circulomics Short-Read Eliminator XL Kit (Circulomics Cat# SKU SS-100-111-01). Out of this size selected DNA 1.5 µg was used as input for two library preparations with the Oxford Nanopore LSK-110 ligation sequencing Kit. Half of each library was loaded onto a PromethION PRO-002 Flowcell (R9.4.1 pores) and sequenced for 24 h. Then the Flowcells were rinsed using the Oxford Nanopore Wash Kit WSH-003 and the second part of each library was loaded and sequencing continued for another 48 h. ONT Sequencing was performed on an Oxford Nanopore P24 PromethION at the Forschungszentrum Jülich using MinKNOW version 21.02.7 without live basecalling. Basecalling of the Oxford Nanopore read data was done using guppy basecaller version 5.0.11 using the R9.4.1 PromethION high-accuracy model (dna_r9.4.1_450bps_hac_prom).