GSM9384294: IgG_3 (ATAC-seq); Mus musculus; ATAC-seq
Spleen and dLN from influenza infected mice were homogenized, red blood cells lysed with ACK Lysis buffer (150mM NH4Cl, 10mM KHCO3 and 0.5M EDTA (pH 7.3)). Lymphocytes from influenza infected mice were stained for cell sorting. For each sample, 5,000 cells were sorted into culture media and transposition was performed. Briefly, cells were resuspended in 12.5 μl 2x TD Buffer, 2.5 μl Tn5, 2.5 μl 1% Tween-20, 2.5 μl 0.2% Digitonin, and 5 μl H2O and incubated at 37C for 1 hr. Cells were then lysed with the addition of 2 μl 10 mg/ml Proteinase-K, 23 μl Tagmentation Clean-up buffer (326 mM NaCl, 109 mM EDTA, 0.63% SDS), and incubated at 40°C for 30 min. Tagmented DNA was purified and size selected for small fragments using AMPure XP beads (Beckman Coulter, A63881) and PCR amplified (Roche, KK2602) with dual indexing Illumina primers (Illumina, FC-131- 2004) ) to generate a sequencing library. Final libraries were again purified and size selected using AMPureXP beads, quantitated by QuBit (Life Technologies, Q33231).