GSM4705163: MCF-7T-1-IP; Homo sapiens; MeDIP-Seq
Genomic DNA was sonicated to ～200-800bp fragments, and 1 μg of fragmented sample was ligated to Illumina's genomic adapters, following the manufacturer's instructions. ～300-900 bp ligated DNA fragments were further immunoprecipitated by anti-5-Methylcytosine antibody. The enriched DNA was amplified by PCR and purified by AMPure XP beads. DNA samples were fragmented to a size range of ～200-800bp with a Diagenode Bioruptor. About 1 μg of fragmented DNA was prepared for Illumina NovaSeq 6000 sequencing as the following steps: 1) End repair of DNA samples; 2) A single 'A' base was added to the 3' ends; 3) Ligation of Illumina's genomic adapters to DNA fragments; 4) MeDIP to enrich Methylated DNA by anti-5-Methylcytosine antibody; 5) PCR amplification to enrich precipitated fragments; 6) Size selection of ～300-900bp DNA fragments using AMPure XP beads. The completed libraries were quantified by Agilent 2100 Bioanalyzer. The libraries were denatured with 0.1 M NaOH to generate single-stranded DNA molecules, captured on Illumina flow cell, amplified in situ. The libraries were then sequenced on the Illumina NovaSeq 6000 following the NovaSeq 6000 S4 Reagent Kit (300 cycles) protocol.