GSM3905155: Foreskin fibroblasts, Hi-C quiescence rep2; Homo sapiens; Hi-C
1-2 million of cells were fixed with 1% formaldehyde at room temperature for 10 min. Formaldehyde was quenched with glycine for 10min at room temperature. cells were lysed and digested with MboI overnight with rotation. MboI was then inactivated at 62℃ for 20 minutes. After filling in with biotin-14-dCTP, the fragments were ligated at room temperature for 5.5 hours with rotation. Then reversal of crosslink and DNA purification was done. The biotin-labeled DNA was sheared to 300-500bp with Covaris M220 and then pulled down with Dynabeads MyOne Streptavidin C1 (Life Technology). Sequencing library preparation was performed on beads, including end-repair, dATP tailing and adaptor-ligation. ­­9 cycles of PCR amplification were performed with Extaq (Takara). Finally, size selection was done with AMPure XP beads and fragments ranging from 300bp to 500bp were selected. Truseq
GEO Accession
GSM3905155