GSM8676799: AKPS P1 mouse #801_rep1; Mus musculus; ATAC-seq
Sorted cells were spun at 500g for 5 min at 4°C, lysed in ice-cold ATAC resuspension buffer [10 mM tris-HCl (pH 7.4), 10 mM NaCl, and 3 mM MgCl2] containing 0.1% NP-40 (Sigma-Aldrich), 0.1% Tween 20 (Sigma-Aldrich), and 0.01% digitonin (Promega), pipetted up and down three times, and incubated on ice for 3 min. Cells were then washed in the ATAC resuspension buffer containing 0.1% Tween 20. Cell pellets were resuspended and DNA was tagmented using the enzyme and buffer provided in the Nextera Library Prep Kit (Illumina, FC-121-1031). Tagmented DNA was then purified using the MinElute PCR purification kit (Qiagen, 28004). ATAC-seq libraries were generated using the ATAC-Seq Kit (Active Motif, 53150). Libraries were quantified using the Qubit dsDNA HS Assay Kit (Thermo Fisher Scientific) and profiled using the D5000 ScreenTape on TapeStation 4200 (Agilent Technologies). Libraries were pooled and sequenced on an Illumina sequencer to generate 25 million paired-end 50-base pair reads per library