ATAC-Seq analysis of Wt mouse BMDM cells differentiated in the presence of MCSF (1hIL4, rep1)
Cells were scraped and counted to achieve 50k/ml in ice-cold PBS. Cell suspension was further diluted to 25k/ml and nuclei were isolated with ATAC-LB (10mM Tris-HCl pH7.4, 10mM NaCl, 3mM MgCl2, 0.1% IGEPAL). Nuclei from 25k cells were used for tagmentation using Nextera DNA Library Preparation Kit (Illumina) from two biological replicates. After tagmentation DNA was purified with Minelute PCR Purification Kit (Qiagen). Tagmented DNA was amplified with Kapa Hifi Hot Start Kit (Kapa Biosystems) using 16 PCR cycles. Amplified libraries were purified with AMPure XP magnetic beads. The distribution of libraries was assessed with Agilent Bioanalyzer and sequenced on a HiSeq 2500 platform.