GSM5394164: ATAC_pMEF643B_HAT1_KO; Mus musculus; ATAC-seq
Primary MEFs isolated from HAT1+/+ (n= 3) and HAT1-/- (n=3) embryos (5*10^4 cells) were collected, resuspended in cold PBS, and centrifuged for 5min at 500g at 4°C. Next, cells were washed with 1mL cold RSB buffer (10mM Tris-HCl, pH 7.4, 10 mM NaCl, and 3mM MgCl2). Cells were centrifuged at 500g for 10 min at 4°C, resuspended in 250µL of freshly made Lysis Buffer (250µL of RSB buffer and 2.5µL of 10% NP-40), and incubated for 5 minutes on ice. Cells were centrifuged at 500g for 10 min at 4°C to isolate the nuclei. Isolated nuclei were resuspended in 50µL of the transposition mix (22.5µL of H2O, 25µL of the 2X Tagment DNA Buffer, 2.5µL of Tagment DNA Enzyme (both Illumina)) and incubated at 37°C for 30 min. Transposition reactions were purified with the MinElute PCR Purification Kit (Qiagen) and eluted in EB buffer. ATAC-seq libraries were amplified 23 with Nextera primers (Illumina) by PCR and purified with AMPure beads (Beckman Coulter).