GSM8185639: WT_hCNCC_PC_HiC; Homo sapiens; Hi-C
Cells were first fixed with 2% formaldehyde for 10 minutes at ambient temperature, followed by quenching with 0.2M glycine for 5 minutes. Post-fixation, cells were lysed, and endogenous nucleases were inactivated with 0.3% SDS. Chromatin was digested using 100U of HindIII (New England Biolabs), labeled with biotin-14-dCTP (Invitrogen), and then ligated using 50U of T4 DNA ligase (New England Biolabs). After reversing the cross-links, DNA was isolated using the QIAamp DNA Mini Kit (Qiagen), in strict accordance with the manufacturer's instructions. Subsequent steps involved fragmenting the DNA to 300-600 bp, end-repair, A-tailing, and adapter ligation using SureSelect adaptors. Biotinylated fragments were isolated through streptavidin-affinity pull-down and PCR amplified. Promoter-specific Capture Hi-C was performed using the SureSelect XT Library Prep Kit ILM (Agilent Technologies), which included a custom-designed biotinylated RNA bait library and paired-end blockers. Post-capture PCR enrichment was followed by final purification of the libraries with AMPure XP beads (Beckman Coulter). Library quality was evaluated using Bioanalyzer profiles (Agilent Technologies), and high-throughput sequencing was performed on the Illumina NovaSeq 6000 platform. promoter capture HiC