GSM8783777: 100242, 1 Kidney; Rattus norvegicus; RNA-Seq
Nuclei were isolated from archived kidney (same animals for Methylseq) using a central section/biopsy (cortex and medulla were included) collected at 4 weeks of age from HSRA-S (n=3) and HSRA-C (n=4) per 10X Chromium Nuclei Isolation Kit (PN-1000493). Single nuclei samples were processed through the 10X Genomics Chromium Single Cell 3' protocol v3.1 per manufacturers' instruction to capture 5000 cells per sample. Single-cell droplets (GEMs) were produced using the 10X Genomics Chromium Controller and barcoded for sample identification. Final libraries were validated on the QIAxcel DNA High Sensitivity bioanalyzer to visualize the expected insert size, and concentration. The pooled 10X library (n=7 samples) was subsequently sequenced twice on the Illumina NextSeq 2000 per 10X Genomics specifications (28x91 cycles) using a P3 100-cycle Kit. Our snRNAseq approach captured an average of 5,293 cells per sample, with an average of 55,945 reads per cell, detecting a median of 1,180 genes per cell.