GSM8445035: GLUP_TEAD4KO_1; Homo sapiens; OTHER
Briefly, about 1 x 105 cells were suspended in 100 μl of wash buffer and incubated with concanavalin A-coated magnetic beads for 1 hour to tether the cells onto the beads. The bead-bound cells were then resuspended in 50 μl of antibody buffer with 1 μg indicated antibody at 4°C with slow rotation overnight. Subsequently, the bead-bound cells were treated with 50 μl of secondary antibody diluted with the dig wash buffer for another hour at RT. After being washed with the dig wash buffer three times to remove unbound antibodies, the cells were further incubated with the hyperactive Pa-Th5 transposase adaptor complex to digest host genome and obtain the fragmented DNA. The fragmented DNA was purified using the phenol-chloroform-isoamyl alcohol extraction and ethanol precipitation. The libraries were amplified by mixing 15 μl the DNA with 5 μl of a universal N5XX and uniquely barcoded N701 primer  followed 15 cycles of 98°C for 10 s, 60°C for 5 s; 72°C for 1 min and hold at 4°C. To purify the PCR products, 1.2x volumes of VAHTS DNA Clean Beads  were added and incubated at RT for 10min.