GSM9111224: αTC1 cells, DMSO, 7 days, biol rep 2.; Mus musculus; ATAC-seq
Nuclei were extracted using nuclei permeabilization buffer and 50,000 nuclei were mixed with tagmentation buffer for tagmentation. 1μL Tn5 transposase (from Yu Miao Lab) was added by pipetting 5 times and then incubated at 37 °C for 60 min in a thermomixer with 500 rpm. Directly following the incubation, sample were purified using the DNA Clean & Concentrator®-5 kit (Zymo research D4014). Fragments were amplified with 2 × NEBNext® Ultra™ II Q5® Master Mix (Biolabs M0544S) and 1 μM primers. PCR conditions were as follows: 72 °C for 5 min; 98 °C for 30 s; and thermocycling at 98 °C for 10 s, 63 °C for 30 s and 72 °C for 1 min for a total number of 14 cycles. Amplified libraries were purified using the DNA Clean & Concentrator®-5 kit. Size selection of libraries were followed using VAHTS DNA Clean Beads (1.5× sample volume; Vazyme N411). Final libraries were quantified using Equalbit dsDNA HS Assay Kit (Vazyme EQ111) and sent to TIANGEN for sequencing.