GSM7664998: HSC_old_CD150_low_ATAC_rep3; Mus musculus; ATAC-seq
About 500 ~ 1000 cells were sorted by FACS to 10 µl tagmentation buffer (33 mM Tris–Acetate, 66 mM K-Acetate, 10 mM Mg-Acetate, 16% Dimethylformamide, and 0.02% Digitonin). 0.5 µl Tn5-Adapter complex (Diagenode) was added after FACS sorting and incubated at 37°C for 30 min. Then, the tagmentation reaction was stopped by adding 10 µl stop buffer (100 mM Tris-HCl pH8.0, 100 mM NaCl, 0.4% SDS, 40 µg/ ml Proteinase K) and the samples were incubated at 55°C overnight to release the tagmentation fragments. SDS was then quenched by adding 5 µl 25% Tween-20 and incubated on ice for 10 min. Next, 25 µl samples were mixed with 30 µl NEBNext High- Fidelity 2X PCR Master Mix (NEB), 2.5 µl P5 primer (10 µM) and 2.5 µl P7 primer (10 µM) for PCR amplification. The PCR products were then purified with SPRI beads with 1:1.6 ratio (96 µl beads) before sequencing.