title: GSM5171873: Day10_HA_H3K27ac_rep2; Homo sapiens; OTHER
library_construction_protocol: 500,000 cells per replicate were pelleted at 300 g for 5 minutes and resuspended in 500 µL 1% methanol-free formaldehyde (Thermo Fisher). After 10 minutes room temperature incubation with rotation, cross-linking was quenched with glycine to a final concentration of 125 mM.  Cross-linked cells were washed in PBS and pellets stored at -80 °C until subsequent next steps. Pellets were resuspended in 500 µL cold Hi-C lysis buffer (10 mM Tris-HCl pH 7.5, 10 mM NaCl, 0.2% NP-40, 1X protease inhibitor (Roche)) and rotated for 30 minutes at 4 °C. Samples were centrifuged at 2500 g for 5 minutes at 4 °C to pellet nuclei. See Mumbach et al (2016, PMID: 27643841) for library construction protocol.  H3K27ac enrichment of 3C libraries.