GSM8460891: F121, GFP+/mCh+, SL2i, CTCF; Mus musculus; ChIP-Seq
For chromatin preparation, 10x10^6 cells (per condition) were cross-linked with a final concentration of 1% formaldehyde for 10 min. The cross-linking reaction was quenched with 2.0 M glycine (0.2 M final glycine concentration). The cross-linked cells were then lysed and sonicated to obtain approximately 300 bp chromatin fragments with a Bioruptor Plus sonication device (Diagenode). For ChIP assays, antibodies were first coupled with Protein G dynabeads (10003D, Thermo Fisher Scientific), and the sonicated chromatin was then incubated overnight at 4○C. After incubation, the beads were washed, the captured chromatin was eluted and cross-linking was reversed. The released DNA fragments were purified with the MinElute PCR Purification kit (28004, Qiagen). The ChIP experiment was performed with the following antibody: CTCF (07-729, Merck Millipore; 5 µL per ChIP). The purified DNA fragments were prepared according to the protocol of the KAPA LTP Library Preparation kit (KR0453, Roche) before sequencing. All ChIP-seq libraries were sequenced with single-end 150 bp cycle mode on an Illumina NextSeq 550 Mid output