title: GSM8285250: K27me3 ALDH plus rep2; Homo sapiens; OTHER
library_construction_protocol: The CUT&Tag sequencing was performed with the CUT&Tag-IT Assay Kit (Active motif, 53160) by following the manufacturer's protocol. 105 OVCAR5 ALDH+ CSCs and OVCAR5 ALDH- non-CSCs were FACS sorted and incubated with Concanavalin A beads for binding. Cells were then incubated with a primary antibody targeting H3K27me3 (Active motif, 39155), H3K4me3 (Abcam, ab8580), and H3K27ac (Active Motif, #39034) overnight at 4°C. Samples were then incubated with secondary antibody, followed by CUT&Tag-IT Assembled pA-Tn5 Transposomes. After tagmentation at 37°C for 1 hour, DNA was purified and cleaned up by SPRI beads. DNA was then amplified with a combination of i7 and i5 Indexed primers for library preparation.