title: GSM6165962: HAP1 SETDB1 KO cells, MBD-seq; Homo sapiens; MBD-Seq
library_construction_protocol: Genomic DNAs (gDNAs) were extracted from the wild-type and SETDB1 KO HAP1 cells using DNeasy Blood & Tissue Kit (Qiagen) according to the manufacturer's instruction, and sonicated using Bioruptor Pico (Diagenode) and purified using with AMPure XP beads (Beckman). Illumina sequencing libraries were generated using the enriched methylated DNA fragments. All the captured DNAs were end-repaired by reacting with T4 DNA polymerase, Klenow, and T4 polynucleotide kinase in the presence of dNTP at 20°C for 2 h, and then 3′-end adenylated using Klenow fragment at 30°C for 2 h. Finally, the end-repaired DNA fragments were ligated with NEBNext adapters (New England Biolabs) and were enriched by 18 cycles of PCR.