title: GSM5258490: H1KD_T47D_HiC_3; Homo sapiens; Hi-C
library_construction_protocol: For the in situ Hi-C, adherent cells were crosslinked with 1% formaldehyde in PBS for 10 min at room temperature. Glycine 0.125M was added for 5 min at room temperature and for 15 min at 4ºC to stop the crosslink reaction. Before permeabilization, cells were treated for 5 min with trypsin. Nuclei digestion was performed with 400 units of MboI restriction enzyme. The ends of the restriction fragments were labeled using biotinylated nucleotides and ligated with T4 DNA ligase. After reversal of crosslinks, DNA was purifed and sheared (Diagenode BioruptorPico) to obtain fragments of around 300-500 bp and ligation junctions were pull-down with streptavidin beads. Hi-C libraries were finally amplified, controlled for quality, and prepared for sequencing using standard Illumina protocols.