GSM5942318: C23; Homo sapiens; ATAC-seq
20 000 to 50 000 sorted CD14+ monocytes were collected in 1mL of RPMI media supplemented with 10% FCS at 4°C.Cells were centrifuged, then cell pellets were resuspended in 50 μL of lysis buffer (Tris HCl 10 mM, NaCl 10 mM, MgCl2 3 mM, Igepal 0,1%) and centrifuged (500 g) for 10 minutes at 4°C. Supernatant was discarded and nuclei were resuspended in 50 μL of reaction buffer (Tn5 transposase 2,5 μL, TD buffer 22,5 μL and 25 μL H2O – Nextera DNA sample preparation kit, Illumina). Tagmentation was performed for 30 minutes at 37°C. DNA was purified using the MinElute purification kit (QIAGEN). Purified DNA was amplified and indexed by PCR using NEBNext High-Fidelity 2× PCR Master Mix (New England Biolabs) with 10-12 cycles. Llibraries were purified using MinElute PCR Purification Kit (Qiagen) and quality controlled using a Bioanalyzer High-Sensitivity DNA Analysis kit (Agilent).