title: GSM8010735: Treatment group; Mus musculus; RNA-Seq
library_construction_protocol: Kidney tissue was minced and cells were dissociated using enzyme mixture (2.5 ml RPMI (no serum), 2 mg/ml collagen protease, 2 mg/ml dispertase and 0.05 mg/ml DNA enzyme I). Single cell suspension was stained in FACS buffer (PBS, 1% BSA, sc-46656, Santa Cruz biotechnology) antibody with fluorescent dye-labeled CD10 (0.05% sodium azide) and sorted with Aria sorp and cell sorter (BD Biosciences, USA). Cell debris was excluded by FSC and SSC parameters, cell adhesion by SSC or FSC W and H parameters, and dead cells by 7-AAD and calcein AM to select cells with good viability. The CD 10-positive cells were then sorted. CD10 + cells and Cd 10-cells were separately sorted from kidney samples and mixed at a 1:2 ratio. The 10X Genomics 3' transcriptome uses short read long sequencing and microfluidic technology for simultaneous transcriptome expression profiling of 500-10,000 cells per sample. The cDNA digestion was first broken into fragments of approximately 200 to 300 bp, plus the traditional second-generation sequencing process of sequencing connector P5 and sequencing primer R1, and finally amplified by PCR to obtain the DNA library. Library sequencing was performed at high throughput using the two-end sequencing mode of the Illumina sequencing platform. At the Read 1 end, 16 bp of barcode information and 10 bp of UMI information were used to determine cells and expression quantification; at the Read 2 end, the cDNA fragment was used to identify the genes corresponding to the mRNA.