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scATAC-seq libraries were constructed by following the paper of Chen, X., et al. (2018). A rapid and robust method for single cell chromatin accessibility profiling. Nature communications, 9(1), 1-9. Briefly, 50,000 cells were cen- trifuged down at 500×g, 4 °C, 5 min. Cell pellets were resuspended in 50 μl tagmentation mix (33 mM Tris-acetate, pH 7.8, 66 mM potassium acetate, 10 mM magnesium acetate, 16% dimethylformamide (DMF), 0.01% digitonin and 5 μl of Tn5. The tagmentation reaction was done at 800 rpm, 37 °C, 30 min. The reaction was then stopped by adding equal volume (50 μl) of tag- mentation stop buffer (10 mM Tris-HCl, pH 8.0, 20 mM EDTA, pH 8.0) and left on ice for 10 min. A volume of 200 μl 1X DPBS with 0.5% BSA was added and the nuclei suspension was transferred to a FACS tube. DAPI (Thermo Fisher 62248) was added at a final concentration of 1 μg/μl to stain the nuclei. |
OTHER |
GENOMIC SINGLE CELL |
PCR |
PAIRED
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