GSM8374609: ATACseq-SKO_1_ATAC; Mus musculus; ATAC-seq
Cells were collected by incubating in trypsin for 5 min at room temperature and subsequent centrifugation at 592g for 5 min at 4 °C. Fifty-thousand cells were used for tagmentation by incubating in 50 μl of 1× THS-seq buffer (25 μl 2× THS buffer (66 mM Tris acetate, pH 7.8, 132 mM potassium acetate, 20 mM magnesium acetate and 32% dimethylformamide), 5 μl 10× Digitonin, 2 μl Illumina-TDE1) for 20 minutes at 37 °C. To stop the tagmentation reaction, an equal volume of 2× Tagmentation Stop Buffer (10 mM Tris-HCl (pH 8.0), 20 mM EDTA (pH 8.0)) was added to the reaction and incubated for 10 min on ice. For cell lysis, an equal volume of 2× lysis buffer (100 mM Tris-HCl (pH 8.0), 100 mM NaCl, 40 μg ml−1 proteinase K, 0.4% SDS) was added to the tagmentation mix and incubated at 65 °C for 15 min. Library construction was performed using standard Illumina protocols using a Nextera Library Prep Kit.