GSM5426088: BJAB-EBNA2-ATAC-Rep2; Homo sapiens; ATAC-seq
For HEK293T ATAC-seq, transient transfection of EBNA2 and vehicle plasmid into BJAB cell respectively, collect positive cells by fluorescence-activated cell sorting (FACS) after 48 hours. To profile open chromatin, a total of 5×104 cell pellets were washed once with cold PBS, cells lysed on ice for 3 minutes in 50 μL ice-cold Lysis Buffer (10mM Tris pH 7.4, 10mM NaCl, 3mM MgCl2, 0.1% NP-40, 0.1% TWEEN 20, 0.01% Digitonin in DEPC H2O), resuspended in 1mL ice-cold RBS-Wash (10mM Tris pH 7.4, 10mM NaCl, 3mM MgCl2, 0.1% TWEEN 20) and pelleted at 4°C at 500 × g for 10 minutes. Tagmentation was performed in 1 × Tagmentation Buffer (10mM Tris pH 7.4, 5mM MgCl2, 10% DMF, 33% PBS, 0.1% TWEEN 20, 0.01% Digitonin) using 100nM Tn5 Transposase for 30 minutes at 37°C. Tagmentation was inactivated with the addition of 5 volumes SDS Lysis Buffer (100mM Tris pH 7.4, 50mM NaCl, 10mM EDTA, 0.5% SDS in H2O) and 100μg Proteinase K (Invitrogen, 25530015, USA) for 30 minutes at 55°C. Centrifuge at 500 × g to take the supernatant, DNA was size selected and purified using QIAquick PCR Purification Kit (QIAGEN, 28106, GERMANY) according to the manufacturer's instructions. ATAC library was prepared according to Vazyme's instructions accompanying the TruePrep DNA Library Prep Kit V2 for Illumina (#TD501).