GSM7846560: Obese CD8 TIL, scRNA-seq; Mus musculus; RNA-Seq
CD8+ TIL were enriched from tumors using CD8+ MicroBeads and separated on LS columns using a QuadroMACS Cell Separator. Enriched TIL were then sorted to at least 99% purity based on CD8 staining using a BD FACSAria III (BD Biosciences). Single-cell RNA libraries were prepared using the Chromium Single-Cell 3' Library and Gel Bead Kit. GEM-RT was performed in a T100 96-Well Thermal cycler according to 10X protocol. The GEMs were then broken, and single-stranded cDNA was cleaned up with DynaBeads MyOne Silane Beads. Barcoded, full-length cDNA was amplified using the T100 96-Well Thermal Cycler according to 10X protocol and stored at -20°C. Amplified cDNA product was cleaned up with the SPRIselect Reagent Kit. 3' gene-expression libraries were constructed using the reagents from the Chromium Single-Cell 3' Library Construction Kit.