GSM8773928: CAF-1 OFF 48 h scRNA-seq; Mus musculus; RNA-Seq
Cell suspension was collected and enriched for live cells by removing dead cells (Dead Cell Removal Kit, Miltenyi Biotec). Cell concentration and viablity were determined for each sample via automated cell counter (Countness II, Thermo Fisher Scientific) with Trypan blue staining (Trypan Blue solution, Sigma-Aldrich). The cell concentration was adjusted with cells resuspended at 1,000 cells per uL in DPBS (DPBS, Thermo Fisher Scientific) with 0.05% BSA (Ultrapure BSA, Thermo Fisher Scientific) for 10x Chromium GEM-X Single Cell 3' v4 scRNA-seq. Gel Bead-in-emulsion (GEM) generation and barcoding was peformed by University of California, Irvine Genomics, Research, and Technology Hub following 10X Chromium GEM-X Single Cell 3' Reagent Kits v4 User Guide. Single live cells suspension with the concentration of 1,000 cells per uL were loaded onto 10X Genomics Chromium GEM-X Chips according to manufactor recommendations. Reverse transcription and library construction were performed with 10X Chromium GEM-X Single Cell 3' Reagent Kits v4 following manufacter user guide. Sequencing was performed on Illumina NovaSeq X Plus.