GSM1908913: E14_SEbait1; Mus musculus; OTHER
4C-seq experiments and analysis was performed as described previously1. Briefly, 5 million cells were cross-linked with 2% formaldehyde for 10 min at room temperature (RT) and quenched by adding 125 mM Glycine with 5 min additional incubation at RT. Cells were lysed and nuclei were isolated and digested with Csp6I (Thermo Scientific) over night (o/n). Enzyme was inactivated by heat at 65 °C for 20 min. The digested chromatin was subjected for ligation for 16 h with T4 ligase (Life Technologies). DNA was then purified with phenol/chloroform extraction and ethanol precipitation before the second digestion with NlaIII (NEB) at 37 °C o/n.  After enzyme inactivation, a second ligation was performed at 16 °C for 4 h and DNA was purified, of which 4.8 μg in total was used for PCR amplification using 4 different pairs of primers TruSeq