GSM8375162: Runx2-OE SCs cJun-1 cut tag; Rattus norvegicus; OTHER
1×10^6 cells were washed twice gently with cell wash buffer. A total of 10μL Concanavalin A coated magnetic beads were added per sample and incubated at room temperature for 10 minutes. Unbound supernatant was removed. Bead-bound cells were resuspended with Dig wash buffer and incubated with a 1:50 dilution of primary antibody (Runx2、CTCF、cJun、C/EBP-β) on a rotating platform overnight at 4℃. The primary antibody was removed using a magnet stand. The secondary antibody was diluted 1:100 in Dig wash buffer and cells were incubated at room temperature for 60 min. Cells were washed using the magnet stand 2-3 times in the Dig wash buffer. A 1:100 dilution of the pA-Tn5 adapter complex was prepared in Dig-med buffer and incubated with cells at room temperature for 1h. Cells were washed 2-3 times for 5 min in 1 mL Dig-med buffer. Then cells were resuspended in Tagmentation buffer and incubated at 37 °C for 1 h. DNA was purified using phenol-chloroform-isoamyl alcohol extraction and ethanol precipitation. DNA fragments ranging from 250 to 350 bp were kept to execute library preparation. The library products were sequenced by using the Illumina Novaseq 6000 system.