GSM8565047: WT_scRNAseq; Mus musculus; RNA-Seq
Tumor tissues from B16-IRF7-WT/MUT tumor-bearing mice were prepared into a single-cell suspension using collagenase II (1 mg/ml) and collagenase IV (2 mg/ml) at 37 °C for 30 minutes. Red blood cells were removed using lysis buffer, and the suspension was resuspended in flow staining buffer. Live immune cells (CD45 positive and 7-AAD negative) were isolated by FACS and resuspended in sample buffer for single-cell capture. These live CD45+ immune cells were then captured using the BD Rhapsody™ system and lysed. Single-cell suspension was added to the 10x Chromium chip following the instructions provided for the 10X Genomics Chromium Single-Cell 3' Kit (V3), with the goal of capturing 8,000 cells. cDNA amplification and library construction were performed according to standard protocols. Libraries were sequenced on an Illumina NovaSeq 6000 sequencing system (paired-end sequencing, 150 bp) at a minimum depth of 20,000 reads per cell.