GSM7719541: Right kidney, Sham operated, 24h, rep1, snRNAseq; Mus musculus; RNA-Seq
Nuclei were isolated from a section of the kidney as per published protocol (Leiz et al., 2021). Briefly, kidney section was minced and homogenized in EZ lysis buffer. The homogenate was filtered through a 40 µm cell strainer and centrifuged to pellet the nuclei. The nuclei pellet was resuspended and layered onto a sucrose gradient and centrifuged to separate and remove any other cellular components and debris. The purified nuclei were resuspended in an appropriate volume of resuspension buffer. Single nuclei were then sorted based on DAPI fluorescence using a flow cytometer. The sorted nuclei were counted using a CellDrop FL Automated Cell Counter (DeNovix) to determine the concentration. The single nuclei suspension was loaded onto a Chromium Controller (10x Genomics) for single nuclei encapsulation, barcoding and cDNA synthesis. Library preparation using the Chromium Single Cell 3' Reagent Kit v3 (10x Genomics) following the manufacturer's instructions. The prepared libraries were sequenced on an MGI FCL sequencer.