title: GSM2170035: 4C-seq Tnfaip3 in ESC; Mus musculus; OTHER
library_construction_protocol: cells were fixed using 1% formaldehyde and stopped by adding Glycine (0.125M). After lysis, soluble chromatin was subjected to restriction endonuclease digestion: First digestion with HindII, second digestion with DpnII. Intra-molecular ligations were performed using T4 DNA ligase (NEB), reverse crosslinked and purified. The purified chromatin was subjected to RNase digestion and further purified. PCR reactions were performed with a first primer designed on the viewpoint and a second outer primer designed beside the DpnII site. Both primers contained illumina sequencing adapters and Barcodes for multiplexing. Approximately 16 reactions were performed for each viewpoint to generate complex libraries.