GSM6943475: FPD_52.3; Homo sapiens; ATAC-seq
Neutrophils from human patient/control peripheral blood were isolated using the EasyStep Human Neutrophil Isolation Kit (Stem Cell). 50,000 cells were collected and washed with cold PBS. Cell pellets were resuspended in 50 µl of lysis buffer (10 mM Tris-HCl, pH 7.4, 10 mM NaCl, 3 mM MgCl2 and 0.1% Tween-20, 0.1% NP-40, 0.01% Digitonin, 1% BSA) and incubated on ice for 4 min. Lysis was halted with the addition of 50 uL of wash buffer (10 mM Tris-HCl, pH 7.4, 10 mM NaCl, 3 mM MgCl2 and 0.1% Tween-20, 0.1%, 1% BSA). Samples were centrifuged at  500 x g, 4 °C for 5 min. Nuclei pellets were resuspended in 50 µl of wash buffer and centrifuged at  500 x g, 4 °C for 5 min.  Nuclei pellets were resuspended in 50 µl of transposition reaction mix (1× Tagment DNA Buffer, 2.5 µl of Tagment DNA Enzyme 1) and incubated for 30 min at 37 °C Libraries were purified using SPRI-Select bead purification and the concentrations were measured using both Qubit and KAPA qPCR. 2100 Bioanalyzer was used to check the quality of libraries. Libraries were sequenced on the Illumina NextSeq 250, with 75-bp paired-end reads.