title: GSM7797137: Enl-WT, snATAC-seq; Mus musculus; ATAC-seq
library_construction_protocol: Kidneys from E15.5 Enl-WT, Enl-T1, and Enl-T1 with TDI-11055 treatment embryos were harvested and incubated with digestion solution containing Enzyme P, Enzyme D, and Enzyme A from the Multi Tissue Dissociation Kit 2 (Miltenyi Biotec, #130-110-203) in a gentleMACS C tube. The tissues were digested on the gentleMACS Octo Dissociator with Heaters run program 37C_Multi_E. The samples were resuspended and filtered through 70 µm strainer, followed by one wash with 15 ml PBS. After centrifugation at 300×g for 10 min, the cell pellet was incubated with 10 volumes of 1X Red blood cell lysis solution (Miltenyi Biotec, #130-094-183) for 2 min at RT. Then, the samples were centrifuged at 300×g for 5 min at RT and resuspended with PBS for further steps. Cell number and viability were measured on a Countess AutoCounter (Invitrogen, #C10227). snATAC-seq was performed with Chromium Next GEM Single Cell ATAC Reagent Kit V2 (10x Genomics, #PN-1000406) in accordance with the manufacturer's instructions (10x Genomics, CG000496 RevA). Briefly, 10,000 nuclei were loaded and incubated in the transposition buffer by targeting 6,000 nuclei recovery. The GEM was generated on Chromium Next GEM Chip H in the Chromium Controller. After the GEM incubation cleanup, the library was constructed according to the manufacturer's manual. Qubit was used for DNA concentration measurement and Agilent Bioanalyzer High Sensitivity DNA kit was used for library quality control according to the manufacturer's manual.