title: GSM6604376: ATAC-seq_mixed H7 cells,positive,Week 5; Homo sapiens; ATAC-seq
library_construction_protocol: For the ATAC sequencing, the mixed organoids were dissociated into single cells with Papain dissociation kit. Then, the dissociated cells were divided into eGFP-positive and eGFP-negative portions using a BD FACS Aria, and 50,000 of sorted cells were lysed with 50 ul of nuclei isolation buffer (10mM Tris-HCl (pH7.4), 10 mM NaCl, 3 mM MgCl2, 0.1% Tween-20, 0.1% NP-40, 0.01% Digitonin, 1% BSA, 1 mM DTT) for 5 min on ice. The 500 ul of chilled Wash Buffer (10mM Tris-HCl (pH7.4), 10 mM NaCl, 3 mM MgCl2, 0.1% Tween-20, 1% BSA, 1 mM DTT) were added to lysed cells, and cells were centrifuged at 500 x g for 5 min at 4°C. The supernatant was removed, and transposition was performed using Tn5 (illumina). The fragmented DNA by Tn5 was purified using MinElute reaction Cleanup kit (Qiagen) and amplified with predesigned primers containing adaptors and barcodes. Next, the ATAC-seq libraries were purified using AMPure XP beads and final libraries were sequenced with a paired end read of 150 bp on the NovaSeq 6000 illumina platform.