title: GSM4314808: H9_P4CNCC_ATACseq_replicate2; Homo sapiens; ATAC-seq
library_construction_protocol: ATAC-seq was performed as described previously (Buenrostro et al., 2013; Corces et al., 2017) Briefly, cells were dissociated and treated with DNaseI (Worthington) and 50,000 viable cells were sorted as DAPI negative. Cells were pelleted at 500 RCF for 5 min at 4°C and resuspended in ATAC-resuspension buffer containing 0.1% NP40, 0.1% Tween­20, and 0.01% Digitonin and incubated on ice for 3 minutes. Following wash-out with cold ATAC-­Resuspension Buffer (RSB, 10 mM Tris­-HCl pH 7.4, 10mM NaCl, 3mM MgCl2 in sterile water) containing 0.1% Tween­20, cells were pelleted and resuspended in 50μL transposition mix (25 μL 2x TD buffer, 2.5 μL transposase (100nM final), 16.5 μL PBS, 0.5 μL 1% digitonin, 0.5 μL 10% Tween­20, 5 μL H2O) and incubated for 30 minutes at 37°C with shaking. The transposition reaction was purified using the Zymo DNA Clean and Concentrator­ Kit, and amplified using PCR primers defined in (Buenrostro et al., 2013). Libraries were purified using the Zymo DNA Clean and Concentrator­ Kit, quantified using the KAPA Library Quantification kit and quality assessed by Bioanalyzer (also used to determine average size for pooling libraries). Samples were sequenced on the HiSeq4000 platform (2x 75bp).
attributes.tag: GEO Accession
attributes.value: GSM4314808