GSM8842274: Retina, si-RORb, scRNA-seq; Mus musculus; RNA-Seq
Retinal cells were washed and resuspended in 0.04% BSA/PBS, then counted using a LUNA-FX7™ Automated Fluorescence Cell Counter (Logos Biosystems). Single-cell RNA-seq libraries were prepared using the 10x Chromium controller and Next GEM Single Cell 3' Reagent v3.1 kits following the 10x Genomics protocol. Briefly, single-cell Gel Bead-in-emulsion (GEM) was generated with a target recovery of 10,000 cells, and mRNA was barcoded and reverse-transcribed within GEM. Barcoded cDNA was amplified by PCR and processed for 3' Gene Expression Library construction, including enzymatic fragmentation, end-repair, A-tailing, adaptor ligation, and index PCR. Libraries were quantified by qPCR (KAPA) and validated using the Agilent 4200 TapeStation (Agilent technologies). Sequencing was performed on an Illumina NovaSeq platform.