GSM5491569: TS600_g11_IgG_ACTseq; Homo sapiens; OTHER
ACT-seq was largely performed according to Carter et al (Carter et al., 2019) as described in detail recently (Liu et al., 2021) on triplicates of TS600 and TS667 NTC and KD2 lines. For CUT&Tag experiments, nuclei isolation was performed according to the published protocol (Narayanan et al. 2020) using a minimum of 105 nuclei. Isolated nuclei were washed and centrifuged at 600 g for 4 minutes. Nuclei were resuspended in 200 ul antibody buffer containing 4 ug MEOX2 antibody or IgG for control and incubated at 4 ºC overnight. bulk library was prepared using 30 μL of 2 x Kapa, 20 μL of bulk DNA, 5 μL of Index 1 (i5 10 μM) and 5 μL of index 2 (i7 10 μM) and run in a thermal cycler for 17 cycles. Concentrations were measured using Nano Drop. QC was performed using D1000 Tapestation. The samples were sequenced on NextSeq 550 (paired-end 75-bp mid-output).