GSM6797229: nephrectomy 2 RNA; Homo sapiens; RNA-Seq
[Single nucleus isolation for multiomic libraries]  For single nucleus multiomic libraries, samples were cut into <2 mm pieces and homogenized using Dounce loose head homogenizers. Samples were then filtered through a 200µm cell strainer and homogenized in the Dounce tight head homogenizers. Samples were incubated on ice for 5 minutes, then filtered through a 40µm cell strainer, centrifuged at 500g for 5 minutes at 4°C. The resuspended pellet was then washed with 4ml of lysis buffer and incubated for 5 minutes at 4°C. Then the sample was centrifuged again and resuspended, then filtered through a 5µm cell strainer. After counting, nuclei suspensions were diluted if needed to target 10,000 nuclei per lane and loaded into a thermal cycler to begin the transposition reaction, following 10X Genomics' protocol. The manufacturer protocol was followed for the completion of library preparation. [Single nucleus isolation for snRNA-seq or snATAC-seq libraries from human adult kidney biopsy samples]  For split single nucleus snRNA-seq and snATAC-seq from biopsies, the second spin step was omitted from the nucleus isolation protocol in order to preserve nuclei. Single nucleus suspensions were loaded into the Chromium Controller following manufacturer instructions in order to target 10,000 nuclei per lane. Lanes that were resequenced were combined with cellranger count into a single output for the entire lane. For kidney samples with enough nuclei, we prepared additional lanes, then combined lanes into a single output for the original kidney with cellranger aggr. 10x chromium 3' RNA v3, ATAC v1.1, and multiome kits used for preparation