GSM3722648: day0_3 ATAC-seq; Canis lupus familiaris; ATAC-seq
Day 0 and 2D cells were collected using cell scraper, and 3D cysts were isolated from m3atrigel with cell recovery solution (Corning). After washing with cold PBS, the cells were dissociated in lysis buffer (1% NP40, 10mM Tris pH7.5, 10mM NaCl, 3mM MgCl2, 1mM EDTA) using syringe. The samples were filtered with miracloth. Crude nuclei were sorted by flow cytometer (Beckman Coulter MoFlo XDP) with DAPI. The nuclei were pelleted and resuspended in the transposase reaction mix (25 μl 2xTagmentation buffer, 2.5 μL transposase and 22.5 μL nuclease-free water), and incubated at 37°C for 30 min. Immediately following transposition, transposed DNA was purified with Qiagen minElute Reaction Cleanup Kit, and amplified using Phusion DNA polymerase (NEB) for 10-13 cycles.