GSM9115340: ATAC, Paused, WT, rep 3; Mus musculus; ATAC-seq
50,000 live cells were collected and resuspended in cold resuspension buffer (RSB: 10 mM Tris-HCl, pH 7.4; 10 mM NaCl; 3 mM MgCl₂) supplemented with 0.1% NP-40, 0.1% Tween-20, and 0.01% digitonin. Following lysis, nuclei were washed in RSB containing 0.1% NP-40 and subjected to tagmentation using the transposition mixture: 25 μL of 2× TD buffer, 2.5 μL of Tn5 transposase (100 nM final concentration), 16.5 μL of 1× PBS, 0.5 μL of 1% digitonin, 0.5 μL of 10% Tween-20, and 5 μL of nuclease-free water. After transposition, DNA was purified using the Zymo DNA Clean & Concentrator-5 Kit. Library amplification was carried out as per protocol (Corces et al., 2017). Amplified libraries were purified using AMPure XP beads with a double-sided size selection to remove both primer dimers and large fragments (>1000 bp). Final libraries were quantified using the Qubit dsDNA High Sensitivity Assay Kit, pooled at equimolar concentrations, and sequenced on a NovaSeq X platform with a 10B flow cell, generating 150 bp paired-end reads.
GEO
GSM9115340_r1