GSM9541285: KO_Ctrl_1; Mus musculus; ATAC-seq
6,000 cells were sorted by FACS into PBS buffer supplemented with 0.5% BSA. For library preparation, cells were lysed and tagmented with 9.5uL lysis and transposition mix buffer (10mM TAPS-NaOH pH 8.5, 5mM MgCl2, 10% DMF, 0.02% digitonin) and 0.5uL Tn5 transposase (Diagenode) at 37 °C for 30 min. Tagmentation was terminated by the direct addition of 10 µL stop buffer (100 mM Tris-HCl pH 8.0, 100 mM NaCl, 0.4% SDS, 40 µg/mL proteinase K), followed by incubation at 55 °C overnight. Subsequently, 5 µL of 25% Tween-20 was added directly to the reaction. Libraries were amplified by PCR for 16 cycles under the following conditions: 72 °C for 5 min; 98 °C for 3 min; followed by cycling at 98 °C for 15 s, 63 °C for 30 s, and 72 °C for 1 min; with a final extension at 72 °C for 5 min. PCR products were purified using a two-step SPRI beads cleanup (1.5× followed by 0.6×). Library concentration was measured using the Qubit dsDNA HS Assay Kit (Invitrogen), and fragment size distribution was assessed using an Agilent FA5400 Fragment Analyzer. Final libraries were sequenced on an Illumina NextSeq™ 1000/2000 platform using a P2 XLEAP-SBS™ Reagent Kit (100 cycles; Cat. No. 20100987).