GSM8520426: Lu134A, ASCL1; Homo sapiens; OTHER
Lu134A cells (1*10^5) were bound to concanavalin-coated magnetic beads (Bangs Laboratories). After permeabilization with digitonin, cells were incubated with primary antibody at room temperature for 2 h and subsequently incubated with secondary antibody for 1 h. Then, CUTANA pAG-Tn5 for CUT&Tag (EpiCypher) was bound to secondary antibody. After incubation with Mg2+ containing solution to cleave the target DNA, the extracted DNA was amplified by PCR. The PCR product was purified using AMPureXP magnetic beads (Beckman Coulter). Sequence libraries were pooled at 5 nM. For non-specific IgG, the entire library was pooled. The pooled library was sequenced on HiSeqX (Illumina) using 150bp paired end reads.