GSM6901079: MCF-10A_ATAC-seq_rep2; Homo sapiens; ATAC-seq
standard ATAC-seq protocol(Nat Methods 2017, 14 (10), 959-962.) ATAC-seq The cells were fixed using 0.5% PFA for 2 min at RT, and then quenched by 0.125 M glycine for 2 min at RT. The fixed cells were washed with wash buffer (20 mM HEPES pH 7.5, 150 mM NaCl, 0.5 mM spermidine) for 3 times and resuspended in wash buffer. Next, 5K-100K cells were immobilized to concanavalin A–coated beads (Novoprotein, Shanghai) with incubation at RT for 10 min. The beads-bound cells were incubated in 50 μl of blocking buffer (wash buffer with 2 mM EDTA, 0.1% BSA, 100 nM random ssDNA and 0.05% digitonin) at RT by rotating for 20 min. Then the blocking buffer was removed. The incubated cells were then resuspended with 100 ul of Tn5 buffer (20 mM HEPES pH 7.5, 300 mM NaCl, 0.5 mM spermidine and 0.01% Digitonin) with 100 nM BAMEA-Tn5 and incubated at room temperature for 0.5 hour by rotating. Then the BAMEA-Tn5 buffer was removed. BAMEA-Tn5-bound cells were washed with 500 ul of Tn5 buffer twice, followed by tagmentation in 25 ul of tagmentation buffer (Tn5 buffer with 10 mM MgCl2) at 37℃ for 0.5 hour. Then 25 uL of 100 nM MEB-Tn5 in 20 mM HEPES pH 7.5 was added to the mixture and further incubated for 0.5 h. After tagmentation, 1 uL of 10% SDS was added and further incubated at 55℃ for 10 min to stop tagmentation and detach Tn5 from DNA. Then the DNA fragments were extracted by streptavidin magnetic beads. Purified genomic DNA from AptaChC-seq were mixed with the universal i5 primer and barcoded i7 primer, and amplified with 5X Ampli PCR Mix (Novoprotein, Shanghai). The library PCR products were cleaned up with DNA Clean Beads (Vazyme, Nanjing). OTHER：AptaChC-seq