GSM8635959: OPC, 5day, BAF155 ChIP; Rattus norvegicus; ChIP-Seq
ChIP-seq sample were prepared on day 1 and day 5 after purification of OPC by addition of 1% formaldehyde for 10 min, followed by quenching using 0.125 M glycine. cell membranes were disrupted with lysis buffer to release the nuclear material. The chromatin was sheared into small fragments, usually ranging from 200 to 1000 base pairs, through ultrasonic fragmentation. This was followed by enriching the protein-DNA complexes using the BAF155 antibody (Cell Signaling Technology, catalog number 11956). The BAF155 antibody was bound to Protein A+G magnetic beads (Magna ChIP™, catalog number 16-663), and the sonicated samples were incubated overnight with the antibody-bound beads. After incubation, unbound material was removed, and DNA fragments were released from the immunoprecipitation complexes. The purified DNA underwent end repair and splice addition, followed by amplification using PCR to generate a sequencing library. Once the library was tested and validated, it was sequenced using the Illumina HiSeq platform.