GSM9588156: XIII_CTL_ChIP_ELOBrepA; Homo sapiens; ChIP-Seq
For ChIPseq, a single cell suspension of control or ELOBKO SU-DIPGXIII+Cas9 cells collected 14 days after sgRNA infection were crosslinked with 1% formadehyde for 10 minutes followed by quenching with glycine. Crosslinked chromatin was sonicated to produce DNA fragments 150 to 2000 bp in length. Soluble chromatin was pre-cleared by adding 150 ul of a 50:50 slurring of pre-washed protein G beads to each tube and incubating at 4°C for 1 hour on a rotator. Exracted chromatin from 2e6 cells was used in each IP with 5 ug of either anti-ELOB (Abcam #ab45297) or with an RNA polymerase II CTD repeat YSPTSPS antibody [8WG16] antibody (Abcam #ab817) Libraries were constructed from 5 ng of input DNA using the NEBNext Ultra II DNA Library Prep Kit for Illumina (E7645, E7103) following the manufacturer's instructions