title: GSM8491160: DMSO_SMARCA4-dTAG_Rep3; Mus musculus; ATAC-seq
library_construction_protocol: For ATAC-seq 60.000 cells were collected and resuspended in 500 µl RSB buffer (10 mM Tris-HCl pH7.4, 10 mM NaCl, 3 mM MgCl2). Cells were pelleted at 500 rcf, at 4°C for 5 min. Supernatant was removed, cells were resuspended in 50 µl RSB buffer containing 0.1% Tween-20 and 0.1% NP40 and incubated on ice for 3 min. Lysis was washed out with 500 µl RSB buffer containing 0.1% Tween-20. Samples were centrifuged for 10 min at 4°C and 500 rcf. Supernatant was removed and pellets resuspended in 50 µl transposition mix (25 µl TD buffer (Diagenode, C01019043-1000), 2.5 µl Tagmentase (Diagenode, C01070012-200), 22.5 µl H2O). Samples were incubated for 30 min at 37°C at 1000 rpm. DNA was cleaned up using the MinElute kit (Qiagen, 28004). Eluted DNA was amplified for 10 cycles, adding Nextera Ad PCR primers. DNA was cleaned up using the MinElute kit (Qiagen, 28004). Library molarity and quality were assessed by Qubit and Tapestation (DNA High sensitivity chip). Libraries were sequenced on a NovaSeq 6000 Illumina sequencerwith paired-end 50 settings.