GSM3075373: LN95_ATAC_seq rep1; Homo sapiens; ATAC-seq
Cells were treated with DNase at a final concentration of 50 U/ml for 30 min. Then cells were washed, trypsinized and resuspended in cold PBS. 50,000 cells were resuspended in cold ATAC-seq resuspension buffer (10 mM Tris-HCl pH 7.4, 10 mM NaCl, and 3 mM MgCl2). Cell nuclei were then prepared by incubation in 50 μl of ATAC-seq resuspendsion buffer containing 0.1% NP40, 0.1% Tween-20, and 0.01% digitonin on ice for 3 min. After centrifugation, nuclei were resuspended in 50 μl of transposition mix (25 μl 2× TD buffer ,2.5 μl Nextera Tn5 transposase (Illuminar), 16.5 μl PBS, 0.5 μl 1% digitonin, 0.5 μl 10% Tween-20, and 5 μl water), and incubated at 37 °C for 30 min in a thermomixer with shaking at 1,000 rpm. Transposed fragments were then purified with a Zymo DNA Clean and Concentrator-5 Kit. All libraries showed sufficient amplification after the 5 pre-amplification cycles and quantified using the KAPA Library Quantification Kit.
GEO Accession
GSM3075373