GSM9350840: WT_U2OS_rep1; Homo sapiens; OTHER
Firstly, cells were trypsinized and genomic DNA was extracted as in Laird et al., 1991 Nucleic Acids Research. In brief, cells were lysed and digested overnight at 37 °C in a buffer containing 0.2% SDS and 100 µg/mL proteinase K. DNA was then precipitated in 100% isopropanol. Libraries were prepared using Oxford Nanopore Technologies Native Barcoding Kit 24 V14 (SQK-NBD114.24) per manufacturer's instructions. Libraries were barcoded, pooled, and loaded onto an Oxford Nanopore Technologies MinION Flow Cell (R10.4.1, FLO-MIN114). Libraries were sequenced for 48 h at a rate of 400-450 bases per second with live basecalling off. Approximately 500 Mb of data per genome were collected, representing ~0.1X genome coverage for an accurate and precise measure of global DNA methylation (per Faulk., 2023 Genome Research).