GSM9014157: hfPSCs_MGT3_DNA methylation; Homo sapiens; Bisulfite-Seq
Genomic DNA was extracted using the phenol-chloroform method. First, cells were lysed with a buffer containing SDS and proteinase K. Then, proteins were removed by phenol-chloroform extraction. Finally, genomic DNA was precipitated with ethanol. Starting with 500ug of genomic DNA for library construction, bisulfite conversion was performed following the protocol of the EZ DNA Methylation-Gold™ Kit (D5006) for DNA methylation analysis to prepare the WGBS library. This workflow primarily includes the following steps: 1) DNA Fragmentation; 2) End Repair; 3) A-Tailing (Adding 'A' to DNA ends); 4) Ligating Methylated Adapters; 5) Size selection via 2% agarose gel electrophoresis and target fragment recovery; 6) C-T conversion (Bisulfite treatment) and purification; 7) PCR amplification to generate the WGBS library