GSM8734557: sh-Hmgb1-3; Mus musculus; ATAC-seq
Cells were lysed using cold lysis buffer (10 mM Tris-HCl, pH 7.4, 10 mM NaCl, 3 mM MgCl₂, and 0.1% IGEPAL CA-630). Nuclei were isolated by centrifugation at 500 × g for 10 minutes at 4°C, and the supernatant was discarded. The nuclei pellet was resuspended in a transposition mix containing 25 μL of 2x TD buffer, 2.5 μL of transposase (S604-02, Vazyme), and 22.5 μL of nuclease-free water, followed by a transposition reaction at 37°C for 30 minutes.After transposition, the sample was purified using the MinElute PCR Purification Kit (28006, Qiagen). Following purification, we amplified library fragments using 1x NEBnext PCR master mix and 1.25 μM of custom Nextera PCR primers, using the following PCR conditions: 72°C for 5 minutes, 98°C for 30 seconds, followed by thermocycling at 98°C for 10 seconds, 63°C for 30 seconds and 72°C for 1 minute, last extension at 72℃ for 5min. The libraries were purified using a Qiagen PCR cleanup kit yielding a final library.