GSM5907292: ATAC_0h1 [ATAC-seq]; Mus musculus; ATAC-seq
About 50,000 spleen cells were lysed with 50 mL of Lysis buffer (10 mM Tris-Cl pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% NP-40, 0.1% Tween-20, protease inhibitor). After centrifuging at 500 g for 10 min at 4oC, pellets (nuclei) were resuspended with 50 ul of transposition mix (Illumina; 1X Tagment DNA buffer, Tn5 Transposase, nuclease-free H2O) and incubated for 30 min at 37 oC in the Thermomixer. Transposed DNA was purified using QIAgen MinElute columns, subsequently amplified with Nextera sequencing primers and NEB high fidelity 2X PCR master mix for 12 cycles (New England Biolabs). PCR-amplified DNA was purified using QIAgen MinElute columns and sequenced using Illumina platform with paired-end reads of 75 bases.