GSM9791192: burb1, untreated, replicate 2; Homo sapiens; OTHER
hiPSCs or hESCs genomic DNA (gDNA) was extracted from isolated nuclei using the Monarch Spin gDNA Extraction Kit (NEB, #T3010S). gDNA concentration was quantified using a Nanodrop. 200ng of gDNA was fragmented to an average size of ~300bp using a Covaris E220 Sonicator with the following parameters: temperature 7°C, peak incident power 140W, duty factor 10%, cycles per burst 200, treatment time 80s. The sheared gDNA was then used to prepare EM-seq libraries using the NEBNext Enzymatic Methyl-seq kit (NEB, #E7120S) following the manufacturer's standard size library protocol. The final sequencing-ready libraries were amplified using the protocol's PCR programme with 4 cycles. Size distribution and yield quantification of the libraries were tested using an Agilent 2100 Bioanalyzer high sensitivity DNA chip (Agilent 5067–4626). Libraries were then multiplexed and sequenced (paired-end 150 bp) on a NovaSeq X Plus Series PE150 (Illumina) to a depth of a minimum 30 million reads per library.