GSM8499388: LCL, Donor 1301, KO; Homo sapiens; ATAC-seq
100,000 cells were spun down at 500g for 5 min at 4°C. Cell pellet was resuspended in 50 µL of fresh ice-cold ATAC-seq Lysis Buffer by pipetting up and down three times and incubated on ice for 3 min. Then, 1 mL of fresh ice-cold ATAC-seq Wash Buffer was added to dilute the lysis reagents and tubes inverted five times to mix. Nuclei were spun down at 500g for 10 min at 4 °C. Nuclei pellets were then resuspended in 50 µL of freshly made Transposition Mix by pipetting up and down six times and reactions were incubated at 37°C for 30 min in a thermomixer with 1,000 rpm mixing. The transposition reaction was terminated by adding 250 µL (five volumes) of DNA Binding Buffer from the DNA Clean and Concentrator-5 Kit and mixed well by inversion. Reactions were kept o/n at -20°C. The next day DNA was purified using the Zymo DNA Clean and Concentrator-5 Kit as per protocol Each sample in the study was assigned to a unique combination of the Adapter 1 (Ad1) and Adapter 2 (Ad2) sequences. Each cleaned-up transposed DNA sample was transferred to a 200 µL PCR tube and mixed with 25 µL of NEBNext Ultra II Q5 2X Master Mix, 2.5 µL of the corresponding Ad1 and 2.5 µL of the corresponding Ad2