title: GSM3731846: ATAC_THP1_24hrPMA_rep1; Homo sapiens; ATAC-seq
library_construction_protocol: THP1 cell suspension volume corresponding to 2E5 cells was pelleted in a 1.5mL tube at 4°C, 500 R.C.F for 5 minutes. Supernatant was aspirated and the pellet resuspended in 150µL cold ATAC lysis buffer (10mM Tris-HCl pH = 7.4, 10mM NaCl, 3mM MgCl2, 0.1% IGEPAL-630). This suspension was agitated by pipetting up/down with a 200µL micropipette tip 8 times. Subsequently, this suspension was pelleted at 4°C, 500 R.C.F for 10 minutes. Supernatant was discarded and pellet immediately resuspended in 190µL transposition reaction mix (10mM Tris-HCl pH=7.5, 5mM MgCl2, 10% Dimethylformamide) by pipetting up/down with a 200µL micropipette tip 3 times. After resuspension 10µL of pre-assembled Tn5 transposome was added. Tube was flicked gently to mix all components and incubated at 37°C, 30 minutes, 700RPM in an Eppendorf Thermomixer. Immediately after the 30-minute incubation ATAC reactions were terminated by adding 1mL Zymo DNA binding buffer and vortexing. This binding buffer and ATAC reaction mixture was purified according to manufacturer instruction in the Zymo DNA Clean and Concentrator-5 kit. Final elution was in 12.5µL nuclease free H2O. Libraries were barcoded and amplified with 8 cycles of PCR amplification
attributes.tag: GEO Accession
attributes.value: GSM3731846