GSM8486609: Cp, fibroblast, wt, HiC, Rep2; Carollia perspicillata; Hi-C
Fibroblast cells or limb cells were dissociated and 1x10^6 or 5x10^5 cells were used for library processing. Cells were fixed in 2% formaldehyde in 10%FCS/PBS. After cell lysis, chromatin was digested with DpnII enzyme (NEB, #R0543), digested ends were marked with biotin-14-dATP (Invitrogen, #19524016) and subsequently ligated. Crosslinking was reversed, DNA precipitated and sheared to a fragment size of 300-600 bp using a S-Series 220 Covaris sonicator. The biotin-containing fragments were pulled down with Dynabeads MyOne Streptavidin T1 beads (Invitrogen, #65602) and ends were repaired using Klenow Fragment DNA polymerase I (NEB, #M0210) and T4 DNA polymerase (NEB, #M0203). Adaptors were added to DNA fragments using NEBNext Multiplex Oligos for Illumina kit (NEB, # E7335S) and sequencing indices were added by PCR using NEBNext Ultra II Q5Master Mix (NEB, #M0544) . Hi-C libraries were generated as three technical replicates and sequenced on a NovaSeq2 as 100bp paired-end reads.