title: HiSeq X Ten paired end sequencing; ATAC-seq in human CD4+T cells, CD8+T cells, CD19+B cells and CD14+ monocytes
design: ATAC-seq in human CD4+T cells, CD8+T cells, CD19+B cells and CD14+ monocytes
library_construction_protocol: human PBMCs were isolated by Ficoll gradient centrifugation from the the whole blood and the different immune cell subsets were isolated with corresponding isolation kit from Miltenyi Biotec. Briefly, 50,000 live cells were freshly isolated and resuspended in lysis buffer (10 mM Tris-HCl pH 7.5, 10 mM NaCl2, 3 mM MgCl2, 0.1% NP40) and pelleted. The cell pellet was resuspended in transposase reaction mix (12.5 μl 2 x TD buffer, 1.25 μL transposase, 11.25 μL nuclease-free water) and incubated at 37°C for 30 min.  Fragmented DNA was purified using Zymo DNA concentrator kit (D4014, Zymo Research) and library was generated by PCR using KAPA HiFi PCR master mix (KK2602, Kapa Biosystems). An appropriate number of PCR cycles was determined by qPCR as described. PCR cleanup of libraries was performed using AMPure beads (A63881, Beckman Coulter) at a 1.2 ratio.