title: GSM7883017: FlcnKD_mESC_Tfe3_1; Mus musculus; OTHER
library_construction_protocol: 5×105 cells were washed once with PBS and resuspended in ice-cold NE1 buffer for 10 min. Cells were fixed in 0.1% formaldehyde and incubated for 2 min and 2.5 M glycine was used to stop cross-linking. Cells were incubated with activated ConA beads for 10 min. The bead-bound cells were resuspended in 100 µL antibody buffer containing appropriate primary antibody. Primary antibody incubation was performed on a rotator at 4 °C overnight. Secondary antibody was diluted in antibody buffer and incubation was performed on a rotator for 1 h. After washing by wash buffer, the cells were incubated with pA-Tn5 (~0.04 uM) on the rotator for 1 h. The cells were resuspended in Tagmentation buffer for 1 h. Tagmentation was stopped by adding 10µL of 0.5M EDTA, 3µL 10% SDS, and 5 µL 10 mg/ml Proteinase K and incubated at 56 °C for 30 min. The DNA was extracted by chloroform. For library amplification, 21 µL DNA was mixed with 2 µL of a universal i5, 2 µL of a uniquely barcoded i7 primer, and 25 µL NEBNext HiFi 2× PCR Master mix. The library was amplified by following cycling conditions: 72 °C for 5 min (gap filling); 98 °C for 45 s; 14 cycles of 98 °C for 15 s and 63 °C for 10 s; final extension at 72 °C for 1 min and hold at 4 °C.