GSM2492380: G4 ESCs_4Cseq_Psite, rep1; Mus musculus; OTHER
4C was done from pools of 60-70 E9.5 embryos or 1-2x10^6 G4 cells per biological replicate. Embryos were minced, treated with collagenase type 2 and passed throw a 70 micrometers cell strainer, and ES cells were harvested and counted. Samples were crosslinked with 2% PFA, frozen with liquid nitrogen and stored at -80º for later processing. After lysis, chromatin was digested with DpnII (New England BioLabs) as primary enzyme and NlaIII (New England BioLabs) as secondary enzyme, and ligated with T4 DNA Ligase (Promega) was used for the two ligation steps. Circular ligated DNA was amplified using specific primers with the Illumina adaptor library included.