GSM8458245: CON-1-2; Homo sapiens; ChIP-Seq
cells were fixed with 1% formaldehyde for 10 minutes. After quenching and cell lysis, chromatin was sonicated using a E220 focused-ultrasonicator (Covans) 1 µg of each fragmented sample was connected to Illumina's genomic adapters with the Genomic DNA Sample Kit (#FC-102-1002, Illumina) following the manufacturer's protocol. Next, DNA fragments of 300–900 bp were denatured at 95°C to form single-stranded DNA, followed by immunoprecipitation using rabbit anti-5hmC. Next, The DNA enrichment was followed by PCR amplification and purification using AMPure XP beads. Quality control (QC) was conducted to evaluate the integrity of the hMeDIP experiments. The DNA fragment mixture library is denatured by 0.1M NaOH to generate single-stranded DNA molecules, which are loaded into the flow cell channels at a concentration of 8 pM, and then amplified in situ. Sequencing was performed using an Illumina NovaSeq 6000, according to the manufacturer's instructions. Sequencing was performed by running 300 cycles using the NovaSeq 6000 S4 Reagent Kit (#2012866, Illumina).