GSM9439290: CTCF-overexpressed AC16, replicate 1, CTCF ChIP; Homo sapiens; ChIP-Seq
Three days after tranfection, cells were fixed with 1% formaldehyde for 10 min at room temperature and quenching with 125 mM glycine for 5 minutes. Cells were lysed, and chromatin was sonicated for 60 min using the Pixul platform (Active Motif), under the following parameters: Pulse (N):50, PRF (kHz): 1.00, Process Time: 60 minutes, Burst Rate (Hz): 20. Chromatin was incubated overnight at 4 °C with Protein G/A Dynabeads (Invitrogen, #10003D) pre-bound to 1.5 µg of anti-CTCF antibody (Active Motifs, #91285). Bead-bound complexes were extensively washed and resuspended in TT buffer (10 mM Tris-HCl, pH 8.0, 0.05% Tween-20). On-bead library preparation was performed using the NEBNext Ultra II DNA Library Prep Kit (NEB, #E7645L). Libraries were barcoded using NEXTFLEX Unique Dual Index Barcodes, PCR-amplified for 14 cycles, with Solexa 1GA/1GB primers, and size-selected (200–500 bp) prior to sequencing on an Illumina NovaSeq SP100 platform
GEO
GSM9439290_r1