GSM5941663: WT_P0; Mus musculus; RNA-Seq
Kidneys were minced with a sterile razor blade. Approximately 65 mg tissue was digested in liberase TM (0.3 mg/mL, Sigma, TM5401119001), hyaluronidase (10 μg/mL, Sigma, H4272), DNaseI (20 μg/mL, Roche, 0471672800) at 37°C for 40 min, followed by incubation with 0.25% trypsin EDTA at 37°C for 30 min. Trypsin was inactivated using 10% fetal bovine serum in PBS. Cells were then resuspended in PBS supplemented with 0.04% bovine serum albumin. After filtration through a 40 μm strainer, cell number and viability were analyzed using Nexcelom Cellometer Auto2000 with AOPI fluorescent staining method. The cDNA and library were generated using the Chromium Next GEM Single Cell 3′ Reagent Kits v3.1 (10X Genomics, PN-1000286) and Single Index Kit T Set A (10X Genomics, PN-1000213) according to the manufacturer's manual.