GSM8995469: D3 WT rep4; Mus musculus; ATAC-seq
50,000 cells were washed in ice cold PBS and lysed in 50 µL of ATAC-seq lysis buffer (10 mM Tris-HCl, pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% NP-40). Following lysis, nuclei were peletted and incubated in 1x transposition buffer (Nextera TD buffer) containing 2.5 μL Nextera Tn5 transposase for 30 min at 37 degrees. Transposed DNA fragments were isolated using QIAGEN MinElute Reaction Cleanup kit and amplified using barcoded primers with Illumina adaptor sequences. The number of cycles used for library amplification was determined using a quantitative PCR side-reaction. Amplified libraries were purified and size selected using Ampure XP beads using consecutive purifications with bead-to-sample ratios of 0.5 and 1.8. After PicoGreen quantification and quality control by Agilent BioAnalyzer, libraries were sequenced on a on a NovaSeq 6000 in a PE100 run using the NovaSeq 6000 S4 Reagent Kit (200 cycles).