title: GSM6380595: Control, 6 months, GEX library; Mus musculus; RNA-Seq
library_construction_protocol: Nuclei were isolated from flash-frozen kidney tissue and processed for single cell multiome ATAC + gene expression sequencing according to the 10X Genomics protocol. All reagents were prepared according to the online protocol. In brief, the tissue of a half kidney was thawed and minced on ice, transferred into NP40 lysis buffer (including Nonidet P40 Substitute, Sigma-Aldrich, 74385), homogenized using a pellet pestle and incubated for 5 minutes on ice. Subsequently, the nuclear suspension was filtered through a 70µm strainer, the flow through was centrifuged, the supernatant removed and the pellet incubated with 1% BSA-PBS + 1U/ul Rnase Inhibitor (Sigma-Aldrich, 3335402001) for 5 minutes on ice. After centrifugation, the pellet was resuspended in 500ul 1% BSA-PBS + 1U/ul Rnase Inhibitor supplemented with 5 ul 7AAD ready made solution (Sigma-Aldrich, SML1633). Nuclei were sorted according to the gating strategy provided in supplemental Figure 3 on a cooled BD Aria I with a 100um nozzle into BSA-PBS + Rnase Inhibitor to achieve a final concentration of 1% BSA-PBS + 1U/ul Rnase Inhibitor. 10X Chromium Single Cell Multiome + Gene Expression protocol (CG000338 Rev E)