GSM6604470: Non-responder-TIL-1-GEX; Mus musculus; RNA-Seq
Tumors and spleens were harvested from responders (n=4) and non-responders (n=4) and single-cell suspensions were prepared. Single-cell suspensions of tumor and spleen from the first cohort (Responder-1, Non-responder-1) were flow sorted for alive CD8 T cells. Single-cell suspensions from the second, third and fourth cohort (Responder-2-4 and Non-responder-2-4) were subjected to EasySep™ Mouse CD8a Positive Selection Kit II (StemCell Technologies, Catalog#18953) according to manufacturer's instructions to purify CD8 T cells. Sorted or purified samples were submitted for single cell capture and library preparation. Cells were loaded into 10× Genomics Chromium Next GEM Chip K (Catalog#1000286) for the 5' captures. Single-cell gene expression libraries were prepared using Chromium Next GEM Single Cell 5' Kit v2, (Catalogue#1000265) according to the manufacturer's instructions. 5' libraries were split in half, one for RNA-seq and another half was enriched for TCR sequencing using the Chromium Single Cell Mouse TCR Amplification Kit (Catalogue#1000254). Samples were sequenced on the Illumina NovaSeq 6000 platform for an estimated read depth of 50,000 reads per cell (5' expression), or 5,000 reads per cell (TCR VDJ).