GSM5582264: Ortho-E_HiC_rep2; Homo sapiens; Hi-C
Cell populations of distinct differentiation stages were isolated by the following cell surface markers: Pro-E cells are CD235a+CD71+CD45lowCD36+CD117highCD105high; Baso-E cells are CD235a+CD71+CD45lowCD36+CD117dimCD105dim; Poly-E cells are CD235a+CD71+CD45lowCD36+CD117-CD105-FSChigh; Ortho-E cells are CD235a+CD71+CD45lowCD36+CD117-CD105-FSClow. Hi-C assay was carried out using the in-situ method as described before with minor modifications17. Half a million FACS-purified primary erythroblasts were used for each reaction. In brief, after fixation with 1% formaldehyde, cells were digested overnight at 37°C using 50U of MboI (NEB, Cat. #R0147). After biotin filling, proximity ligation was carried out overnight at 25 °C with 1600U T4 DNA Ligase (Vazyme, Cat. # N103-01). After reverse-crosslinking, DNA was purified and sheared to 300-500bp fragments using Covaris M220 sonicator. Ligation fragments containing biotin were immobilized on MyOne Streptavidin C1 beads (ThermoFisher Cat. #: 65602), and then library preparation was performed using the DNA library preparation kit (Vazyme, Cat.# ND607) as per the manufacturer's protocol. DNA was then subjected to double-size selection using DNA Clean Beads (Vazyme, Cat. # N411) in order to isolate fragments between 300 and 600bp.