title: GSM8733686: Low-eGFR IgAN3; Homo sapiens; RNA-Seq
library_construction_protocol: Single-cell libraries were prepared using the 10x Genomics Single Cell 3' Library and Gel Bead Kit V3 (Catalog #1000075) and the Chromium Single Cell B Chip Kit (Catalog #1000074). The cell suspension, containing 300-600 viable cells per microliter (quantified using Count Star), was loaded onto the Chromium Single Cell Controller (10x Genomics) to generate single-cell gel beads in emulsion, following the manufacturer's protocol. Briefly, cells were resuspended in PBS with 0.04% BSA, and approximately 16,000 cells were added to each channel, with an expected recovery of around 8,000 cells. Captured cells were lysed, and the released RNA was barcoded during reverse transcription within individual GEMs. Reverse transcription was performed on a Bio-Rad S1000™ Touch Thermal Cycler at 53°C for 45 minutes, followed by 85°C for 5 minutes, with a final hold at 4°C. The cDNA was then generated, amplified, and assessed for quality using the Agilent 4200 (analysis performed by CapitalBio Technology, Beijing). Single-cell RNA-seq libraries were constructed using the Single Cell 3' Library and Gel Bead Kit V3 (10x Genomics), according to the manufacturer's instructions. The libraries were then sequenced on an Illumina NovaSeq 6000 with a sequencing depth of at least 20,000 reads per cell, using a paired-end 150 bp (PE150) read strategy (performed by CapitalBio Technology, Beijing).
identifiers.namespace: GEO
identifiers.id: GSM8733686_r1