GSM6482570: ATAC_WT_Day4_RGFP966 rep2; Mus musculus; ATAC-seq
70000 cultured cells were washed once with PBS and once with cold lysis buffer (10 mM Tris-HCl (pH 7.4), 10 mM NaCl, 3 mM MgCl2, 0.1% NP-40).  The cells were suspended in 50 µl of 1X Reaction Buffer (25 ul of Tagment DNA Buffer, 2.5 ul of Tagment DNA enzyme I, and 22.5 ul of water) (Nextera DNA Library Preparation Kit, Illumina).  Transposase reactions were carried out at 37°C for 30 min, and then DNA was purified using ChIP DNA Clean & Concentrator Kit (Zymo Research). DNA was amplified using the Nextera primer Ad1 and a unique Ad2.n barcoding primer using NEBNext High-Fidelity 2X PCR Master Mix (NEB) for 7 cycles.  The amplified libraries were purified with 2 ul of SpeedBeads (GE Healthcare) in 20% PEG 8000/2.5 M NaCl (Final 13% PEG 8000), eluted with 15 ul of EB (Zymo Research), size selected using PAGE/TBE gel (Invitrogen) for 175-225 bp fragments by gel extraction, and single-end sequenced on HiSeq 4000 (Illumina).