title: GSM5830371: EoKO+EoGFP_rep2 (ChIP-seq); Mus musculus; ChIP-Seq
library_construction_protocol: ChIP seq was done as described previously (Tosic et al., 2019). Briefly, on day 5 of differentiation, EBs were trypsinised and approximately 3x10^7 cells was used per ChIP. Cells were double cross-linked using 2 mM disuccinimidyl glutarate followed by 1% formaldehyde. Chromatin was isolated, sonicated and incubated with 5µg anti-GFP antibody (Abcam, ab290) coupled to ProteinG beads overnight at 4°C. The next day, beads were washed on ice with RIPA buffer and TBS. DNA was eluted, de-crosslinked and purified. Libraries were prepared with NEBNext Ultra II DNA Library Prep Kit for Illumina (NEB, E7645S), size selected using the SPRI select beads (Beckman Coulter) to obtain fragments between 300-600 bp of size.