GSM5839632: ONT_CpG_methyl_PTC_12; Mus musculus; OTHER
For long-read DNA-seq, gDNA was extracted with Qiagen Genomic tip according to manufacturer's specifications. gDNA was fragmented by sonication with a Bioruptor sonicator for 30 cycles of 5 seconds ON – 90 seconds OFF, using low intensity settings. gDNA was then purified with the Zymo DNA Clean & Concentrator™-5 columns (Zymo Research, D4013) and size selected with 0.4X volume of KAPA HyperPure Beads (ROCHE, 08963835001) to exclude small DNA fragments. 1 μg of sheared gDNA was used for library preparation using Ligation Sequencing Kits (Oxford Nanopore Technologies, LSK109 and LSK110) according to the manufacturer's specifications. Samples were sequenced on a MinION sequencer on R9 Flow Cells (Oxford Nanopore Technologies FLO-MIN106D), using the MinKNOW software v19. Flowcells were washed and reloaded with the Flow Cell Wash Kit (Oxford Nanopore Technologies EXP-WSH003) to increase sequencing depth obtained per flowcell. Average read N50 for sequencing runs was 3 kilobases.