GSM8966988: mTH17_ZF7_FICZ_rep2; Mus musculus; ATAC-seq
Nuclei were isolated in ATAC lysis buffer (10 mM Tris-HCl pH 7.5, 10 mM sodium chloride, 3 mM magnesium chloride, 0.1% v/v IGEPAL-CA630, 0.1% v/v Tween-20, 0.01% w/v digitonin) for 3 min on ice. The nuclei were briefly rinsed in wash buffer (10 mM Tris-HCl pH 7.5, 10 mM sodium chloride, 3 mM magnesium chloride, 0.1% v/v Tween-20) before incubating in transposition buffer (10 mM Tris-HCl pH 7.6, 5 mM magnesium chloride, 10% v/v dimethyl formamide, 0.1% v/v Tween-20, 0.01% w/v digitonin) with 100 nM loaded hyperactive Tn5 transposase (Diagenode C01070012) for 30 min at 37 C at 1000 rpm. Transposed DNA was isolated from Qiagen MinElute columns. Libraries were generated using published Illumina-compatible indexed oligonucleotides and 8 cycles of PCR amplification using NEBNext High-Fidelity 2X PCR Master Mix (NEB M0541). Libraries were purified from SPRIselect beads (Beckman Coulter B23317), and 51-bp paired-end sequences were sequenced on a NovaSeq X instrument.