GSM6213139: melanoma cells IP BAHCC1; Homo sapiens; OTHER
GSM6213139
Assays were performed following the manufacturer's instructions (Active motif CUT&Tag-IT™ Assay Kit, #53165, #53160). Briefly, 500 000 cells per condition were used. The cells were washed 2 time before the binding on Concavalin A beads and then incubated overnight with BAHCC1 and HA primary antibodies at the recommended dilution (1:50) or without antibody (negative control). The next day the corresponding secondary antibody, a guinea pig Anti-rabbit antibody was used following a 1:100 dilution in digitonin buffer and incubated at room temperature for 1 hour. The CUT&Tag-IT™ Assembled pA-Tn5 Transposomes were incubated for 1 hour at room temperature before tagmentation. Cells were resuspended in Tagmentation buffer and incubated at 37°C for 1 hour, then the tagmentation process was stopped by addition of EDTA and SDS. Protein digestion was performed by the addition of 80µg/mL of proteinase K and incubated at 55°C for 60 minutes. DNA was retrieved on DNA purification columns provided by the manufacturer. Library preparation and PCR amplification were done using the Kit primers and purified by 2 successive washes with SPRI beads. Libraries were sequenced on an Illumina HiSeq 4000 sequencer as paired-end 100 base reads.