title: GSM8686484: OCI-AML4 DMSO 6hr rep1; Homo sapiens; ATAC-seq
library_name: GSM8686484
library_construction_protocol: Accessible chromatin was assessed using Assay for Transposase-Accessible Chromatin using sequencing (ATAC-seq) as previously described with the Omni-ATAC protocol. Cells were treated with 500nM dTAGV-1 or DMSO for 6 hours and then 50,000 live cells were sorted into PBS with 2% BSA. Cells were washed twice in 150μL and 50μL 1xPBS, resuspended in 50μL ATAC-seq lysis buffer, incubated for 10 min on ice and centrifuged at 400g for 10 min at 4 °C. The pellet was incubated in the transposase reaction mix (25μL 2×TD buffer (Illumina), 2.5μL transposase (Illumina Cat# FC-121-1030) and 22.5μL nuclease-free water) for 30min at 37 °C with gentle agitation. After DNA purification with the Zymo DNA Clean and Concentrator Kit (Zymo, Cat# D4033) libraries were amplified with NEBNext High-Fidelity PCR Master Mix (NEB, Cat# M0541S) using custom Nextera primers. Libraries for sequencing were size selected with Agencourt AMPure XP beads (Beckman Coulter, Cat# A63880). DNA concentration was measured with an Invitrogen Qubit fluorometer (Life Technologies) and Agilent Fragment Analyzer. Libraries were subject to paired-end sequencing with 150 cycles.