GSM7884692
Genomic DNA was isolated from cells using the QIAamp DNA Mini and Blood Mini Kit (Qiagen, 51304). The DNA samples were then assessed for quality and quantity through agarose gel electrophoresis, Nanodrop, and Qubit. After passing quality checks, Firstly, DNA samples were subjected to digestion by the methylation-insensitive restriction enzyme MspI. The resulting DNA fragments were subjected to end repair, A-tailing, and ligation of sequencing adaptors. Gel electrophoresis was then performed to select DNA fragments ranging from 40-220bp in length. Subsequent Bisulfite treatment was carried out using the EZ DNA Methylation Gold Kit from Zymo Research. Finally, PCR amplification was performed to obtain the final DNA library. After library construction, initial quantification was conducted using Qubit 2.0. The library was then diluted to a concentration of 1ng/μl. The insert fragment length of the library was examined using the 5400 Fragment Analyzer System.