title: GSM8860757: neural progenitor cells, C3, Pan troglodytes, 2N, NPC_Rep2; Pan troglodytes; ATAC-seq
library_name: GSM8860757
library_construction_protocol: NPCs at D14 of differentiation or piNs at D28 were lyzed in lysis buffer (10mM Tris-HCl pH7.4, 10mM NaCl, 3mM MgCl2, 0.1% NP-40, 0.1% Tween-20) and collected into conical tubes at a final volume of 500µl lysis buffer. The cell suspension was incubated at 4°C for 10min to release nuclei and then pelleted at 830rcf for 10min. Nuclei were resuspended in 1ml lysis buffer and counted. 50,000 nuclei were then pelleted at 830rcf for 10min and resuspended in 50µl of transposition mix (1X TD buffer [10mM Tris-HCl pH7.6, 5mM MgCl2, 10% dimethyl formamide], 0.33X PBS, 0.001% Digitonin, 0.1% Tween-20, 100nM Tn5 transposase [made in house]). The resuspended nuclei were incubated at 37°C for 30min. DNA was then purified and eluted in 21µl H2O using the DNA Clean and Concentrator-5 Kit (Zymo Research, cat #D4014). 20µl of the eluent was PCR amplified with 1X NEBNext High Fidelity PCR Master Mix (New England Biolabs, cat #M0541L) and primer sequences containing Illumina adapters for 72°C for 5min, 98°C for 30s, and then 5 cycles of 98°C for 10s, 63°C for 30s, and 72°C for 1min.  Libraries were purified using the DNA Clean and Concentrator-5 Kit and quantified with the KAPA Library Quantification Kit (Roche, cat #07960140001).