MicroC of human: HSPC, biological rep2
About 100,000 FACS-purified cells were used for each reaction. In brief, after fixation with 3mM DSG for 30 min followed by 1% formaldehyde for another 10 min, crosslinked cells were permeabilized in ice-cold Micro-C Buffer #1  for 20 minutes. Chromatin from permeabilized cells was digested by 50U micrococcal nuclease at 37C for 10 minutes. After digestion, chromatin fragments were then subjected to dephosphorylation, phosphorylation, end-chewing process. Blunt-end and biotin incorporation was achieved by adding biotin-dATP, biotin-dCTP, dGTP, and dTTP and incubated at 25C for 45 minutes. Chromatin was ligated by T4 DNA ligase at room temperature for 2.5 hours. Unligated ends containing biotin-dNTPs were then removed by exonuclease III in 37C for 15 minutes. After reverse-crosslinking, DNA was purified and gel size-selected for dinucleosomal DNA at 200 to 400 bp. DNA fragments containing biotin were immobilized on MyOne Streptavidin C1 beads  and then library preparation was performed using the DNA library preparation kit (Vazyme, Cat.# ND607) as per the manufacturers protocol. Two biological replicates per condition were sequenced in a NovaSeq 6000 (S4, PE150, Illumina) by Novogene (Beijing).