GSM7401210: Kidney, Lean 3; Rattus norvegicus; RNA-Seq
Rats were perfused with chilled 1x PBS via the left ventricle prior to euthanasia. Single cells were isolated using a previously established protocol. In short, rat kidneys were digested using Miltenyi multi tissue dissociation kit 1 (130-110-2011). Debris was removed using Miltenyi debris removal solution (130-109-398), dead cells were removed using Miltenyi dead cell removal kit (130-090-101). Cell number and viability were analyzed using a Countess AutoCounter (Invitrogen, C10227). 10,000 cells were loaded into the Chromium Controller (10X Genomics, PN-120223) on a Chromium Next GEM chip G Single-Cell Kit (10X Genomics, PN-1000120) to generate single-cell gel beads in the emulsion (GEM) according to the manufacturer's protocol (10X Genomics, PN-1000121). The cDNA and library were made using the Chromium Next GEM Single-Cell 3′ GEM Kit v3.1 (10X Genomics, PN-1000121) and Single Index Kit T Set A (10X Genomics, PN-120262) according to the manufacturer's protocol. Quality control for the libraries was performed using Agilent Bioanalyzer High Sensitivity DNA kit (Agilent Technologies, 5067-4626) for qualitative analysis. Libraries were sequenced on an Illumina Novaseq 6000 system with 2 × 150 paired-end kits using the following read length: 28 bp Read1 for cell barcode and UMI, 8 bp I7 index for sample index and 91 bp Read2 for transcript.