GSM6564308: Formative_ESC_WT_ATAC_rep1; Mus musculus; ATAC-seq
ATAC-seq libraries were generated using the Active Motif commercial kit following the provided protocol with kit components (Active Motif, CAT#53150): Specifically, freshly trypsinized cells were washed with ice cold PBS using 75K cells as input for each sample. Cells were lysed using ice cold ATAC Lysis Buffer and we added 50ul of Tagmentation Master Mix to each sample. We incubated tagmentation reactions at 37C for 30min in a thermomixer set to 800 rpm. We immediately transferred samples to a new tube and performed DNA column purification. After purification we amplified the libraries using the following PCR conditions: 1 cycle at 72C for 5 minutes, 1 cycle at 98C for 30 seconds, and finally 10 cycles of 98C for 10 seconds, 63C for 30 seconds, 72C for 1 minute. Amplified libraries were purified using 1.2x SPRI bead solution. Quality and concentration of libraries were determined by an Agilent 4200 Tapestation with D1000 reagents before pooling for sequencing.