GSM7886619: sorted splenic WT NK cells, rep 2; Mus musculus; ATAC-seq
Nuclei were first extracted for 2 min on ice using nuclei buffer (10 mM Tris-HCl (pH 7.4), 10mM NaCl, 3 mM MgCl2 0.1% Tween-20, 0.1% 755 Nonidet P40 substitute, 0.01% Digitonin, 1% BSA). Nuclei were washed and accessible DNA was tagmented using the Tn5 transposition reaction mix (Illumina Nextera DNA 757 sample preparation kit, FC121-1030; 30 min at 37°C). Tagmented DNA isolated using the MinElute PCR Purification kit (Qiagen, Cat #28004). Following 5 PCR cycles on the thermocycler using a combination of a PCR primer (Ad1_noMX) and an indexed PCR primer, an aliquot was used to assess the number of PCR cycles required for library amplification. The amplified DNA library was subsequently purified using the MinElute PCR Purification kit (Qiagen). Quality and concentration of the library was assessed using Bioanalyser (Agilent).