GSM8379031: NC1235,ATACseq; Homo sapiens; ATAC-seq
For ATAC-seq, 1×105 HCC1954 cells treated with indicated ASOs were harvested and counted. Washed twice with PBS, cells are lysed in Lysis buffer (10 mM Tris-HCl pH 7.5, 10mM NaCl, 3 mM MgCl2, 0.1% NP-40, 0,1% Tween-20, 0.01% Digitonin) on ice for 5 min then centrifuge for 10min at 500 g. Pellets were resuspended in 50 μl 1X TD Buffer (Illumina, 15027865) with 7.5 μl Tn5 Transposase (Illumina, 15027865), 0.01% Digitonin, 0.1% Tween and incubated at 37°C for 30 min. DNA were purified by Monarch PCR & DNA Cleanup Kit (NEB, T1030). Eluted DNA were amplified with NEBNext High-Fidelity 2X PCR Master Mix (NEB, M0541L) and TruePrep Index Kit V2 for Illumina (Vazyme, TD202) for 10 cycles and purified by AMPure XP beads (0.5x-0.7x). The DNA library was prepared using a Next Ultra DNA Library Prep Kit (NEB, E7645) according to the manufacturer's instructions. The libraries were purified with AMPure XP beads (Beckman, A63881) according to the manufacturer 's instructions, and sequenced using Illumina platforms.