GSM9145579: ATAC-SEM-Rep2-3117686_JWH002; Homo sapiens; ATAC-seq
150,000 SEM or K562 cells were lysed by ATAC-RSB+++ on ice for 3 mins. 1mL of ATAC-RSB+ was added to dilute the solution. Nuclei were pellet by centrifugation for 5 minutes, 500g, at 4 degrees. The buffer was discarded while the pellet was resuspended with 10mM N3-Kethoxial in PBS and incubated at 37 degrees, with shaking at 500rpm for 10 minutes. Nuclei were washed by 1mL DPBS and centrifuged for 5 minutes, 500g at 4 degrees to remove the buffer. Following N3-Kethoxial labeling, the transposition reaction was carried out at 37 degrees for 30 minutes. The reaction was terminated by adding 250ul of DNA Binding Buffer from DNA Clean and Concetrator-5 Kit (Zymo). The DNA was eluted by 43ul of 25mM K3BO3. Next, 2.5ul 20mM DBCO-PEG4-biotin (Sigma) was added and incubate the mixture for 1.5hrs, 37 degrees at 500rpm. After the completion fo the Clikc reaction, 2.5ul RNasA for 5 minutes at 37 degrees. The DNA was purified by the DNA by Clean and Concentrator Kit (Zymo). DNA was eluted by 50ul 25mM K3BO3.  The modified DNA was enriched with MyOne Streptavidin C1 beads and incubated for 15 minutes, washed, then purified the DNA with 20ul dH2O. DNA was indexed with 25ul NEBNext HiFi 2x Master Mix (MO543S), 1ul 62.5uM Nextera primer Forward, and 1ul 62.5uM barcoded Nextera primer Reverse. The PCR was caried out for 12 cycles and purified by AMPure XP beads (Beckman Coulter).
GEO
GSM9145579_r1