GSM7412601: PBT_batch5_library1_6donors_CITE; Homo sapiens; RNA-Seq
Tumor-infiltrating T cells were sorted into 1.5 mL tubes containing 500 ul of 1:1 PBS to FBS with 1:100 Recombinant RNAse Inhibitor (Takara Bio 2313B). Following sorting, tubes were filled up to 1.4 mL with PBS and spun at 600xg for 10 minutes. Pellets were resuspended in 35 ul PBS supplemented with 0.04% w/v BSA (Invitrogen) and cells were processed according to manufacturer's recommendations using 10x Genomics' Single Cell Immune Profiling 5'v2 (Dual Index) with Feature Barcoding kit. TCR (VDJ) sequences enriched according to manufacturer's recommendations using 10x Genomics' Single Cell Immune Profiling 5'v2 (Dual Index) with Feature Barcoding kit. HTO (CITE) cDNA was separated from PolyA cDNA through size selection with AMPure XP Purification reagent (Beckman Coulter, A63882) All Libraries were prepared according to manufacturer's recommendations using 10x Genomics' Single Cell Immune Profiling 5'v2 (Dual Index) with Feature Barcoding kit. scRNA-Seq, V(D)J library preparation and Cell Surface Protein library preparation