GSM8141344: DKD, Control, rep2; Mus musculus; RNA-Seq
RNA was harvested with 10X genomics Single-cell protocols. A single-cell suspension of mouse kidney cells was prepared following the 10x Genomics Single-Cell Protocols. Mouse kidney cells, with a concentration of approximately 800-1200/μL, were kept on ice for single-cell analysis. The single-cell suspension was mixed with the master mix and loaded, along with Single-Cell 5' Gel Beads and Partitioning Oil, into the Chromium Next GEM Chip K. The Gel Beads were coated with unique molecular identifiers (UMI), unique primers containing 10x cell barcodes, and oligo poly(dT) sequences. The single-Cell Gel Bead-In-Emulsions (GEM) were generated and barcoded after the GEM generation step using a Chromium Controller (10x Genomics). The barcoded full-length cDNA was constructed via reverse transcription of single cells, followed by incubation using PCR. After encapsulation, cDNAs were selectively purified using Dynabeads Cleanup Mix and then amplified to construct the 5' Gene Expression Dual Index Library.