GSM7081667: Sperm_ATAC_Untreated_Prep3; Mus musculus; ATAC-seq
About 100,000 sperm were on-ice lysed with hypotonic lysis buffer (10 mM Tris-HCl pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% NP40, 0.1% Tween-20, and 0.01% digitonin) for 10 min. Sperm were then resuspended in Tagmentation buffer (10 mM TAPS-NaOH pH 8.5, 5 mM MgCl2, 10% DMF, 0.05% Digitonin) and tagmentation was conducted by adding Tn5 transposase (Diagenode, C01070012) at 1:20 and incubated at 37C for 30min. Reaction was stopped by SDS and proteinase K and incubated at 55°C  overnight. SDS was quenched by tween-20 and library amplification was done with 1x NEBNext High-Fidelity PCR Master Mix containing 0.5 mM indexed primers using the following PCR conditions: 72°C for 5 min; 98°C for 3 min; and 14 cycles at 98°C for 15 s, 63°C for 30 s, and 72°C for 1 min.