title: GSM5981805: 4C-seq_Jurkat-SV_5-aza-dC_TLX3-promoter_rep3; Homo sapiens; OTHER
library_construction_protocol: For 4C-seq analysis using the Illumina NextSeq500, 5 x 106 cells were cross-linked with 2% formaldehyde and nuclei were collected after cell lysis. Fixed chromatin was digested in situ with DpnII followed by in situ proximity ligation using T4 DNA ligase. Chromatin was then reverse cross-linked and genomic DNA was purified using phenol/chloroform extraction followed by ethanol precipitation. Purified DNA was then digested with Csp6I followed by proximity ligation with T4 DNA ligase. 4C templates were purified using QIAquick Spin columns. First round of PCR was performed using reading and non-reading promoter viewpoint primers followed by second round of PCR with primers containing Illumina-compatible adapter sequences.