GSM4224778: 168h_ATAC_rep1; Homo sapiens; ATAC-seq
Cells were collected by centrifugation. Briefly, 5 million cells were harvested and treated with Nextera Tn5 Transposase (Illumina, FC-121-1030) for 45 min at 37°C. Library fragments were amplified using 1x NEBNext High-Fidelity 2x PCR Master Mix (NEB, M0541S) and 1.25uM of custom Nextera PCR primers 1 and 2. PCR amplification was done with 11 cycles, determined by KAPA Real-Time Library Amplication Kit (Peqlab, KK2701) to stop prior to saturation. Then, the samples were purified using Qiagen MinElute PCR Purification Kit (Qiagen, 28004) and with Agencourt AMPure XP beads (Beckman Coulter, A63881) in 3:1 ratio. The libraries were sequenced paired-end (50bp) on a HiSeq2000 instrument