title: GSM8336803: kidney, aged Tfebfl/fl KAP, scRNAseq; Mus musculus; RNA-Seq
library_construction_protocol: All mice were perfused with phosphate-buffered saline (PBS) via the left ventricle to remove blood cells. Afterward, the kidneys were harvested, and the renal cortex was dissected using a razor blade after the renal capsule was removed. Renal cortex samples from two or three mice in each group were combined. These samples were minced with a sterile razor blade and digested in 3.0 ml of Hanks' Balanced Salt Solution (HBSS) containing 250 μg/ml Riberase TH and 100 U/ml DNase I for 30 minutes at 37°C with shaking at 200 rpm. After digestion, the cell suspension was centrifuged at 200  g for 4 minutes. After removing the supernatant, pellets were washed twice with 2.0% bovine serum albumin (BSA) in PBS. Finally, the cell suspension was filtered through a 40 μm cell strainer, and the cells were collected by centrifugation and were resuspended with 2.0% BSA in PBS. Cell viability was investigated by trypan blue staining, and cell concentration was assessed using a hemocytometer. Libraries of the single cells were prepared using Chromium Next GEM Single Cell 3' Library and Gel Bead Kit v3.1 according to the manufacturer's instructions (10x Genomics).