GSM6561110: cGAS-KO, ATRX-KO, Rep1, CUTAC; Homo sapiens; OTHER
For CUT&Tag, tagmentation was performed for 1 h at 37°C in 300-Wash buffer (20 mM HEPES-KOH pH 7.5, 300 mM NaCl, 0.5 mM spermidine) supplemented with 10 mM MgCl2 in a 50 μL volume. For CUTAC, tagmentation was performed for 20 min at 37°C in CUTAC-hex tagmentation buffer (10 mM TAPS, 5 mM MgCl2 10% 1,6-hexanediol). Beads/cells complexes were washed in TAPS wash buffer (10 mM TAPS pH 8.5, 0.2 mM EDTA) using a magnet. Beads/cells were resuspended in 5 μl 0.1% SDS, 10 mL TAPS pH 8.5  solution and incubated  for 1 h  at 58°C to release tagemented DNA. Library from tagmented DNA was performed by adding 15 μL of 0.67% Triton-X100, and 2.5 μL of 10 μM barcoded i5 (v2_Ad1.1 to 1.8) and i7 (v2_Ad2.1 to 2.12) primer solutions (described in (Buenrostro et al., 2015)). A  unique barcode combination was used per sample. Tubes were chilled, and 25 μL of NEBNext 2x Master mix (NEB, M0541L) was added. Using a thermocycler with a ramping of 2°C/sec, PCR reaction was performed as follows: 58°C for 5 min; 72°C for 5 min; 98°C for 30 s, 14 cycles (98°C for 10 s; 60°C for 10 s); 72°C for 1 min. PCR clean-up was performed by addition of 65 μL AMPure XP bead slurry following the manufacturer's instructions (Beckman Coulter #A63881). Fianlly, DNA was eluted with 22 μL 1 mM Tris-HCl pH 8.