title: GSM5516515: 4267_TetPos_Lung_P1; Mus musculus; RNA-Seq
library_construction_protocol: 10x: H-2Kb/SIINFEKL specific CD8+ T cells from KrasG12D+/-  Trp53-/- lung adenocarcinoma bearing lungs at different timepoints were sorted and loaded into the 10X Chromium Controller using the Chromium Single Cell 5' Library and Gel Bead Kit (10X Genomics) Smart-Seq2: H-2Kb/SIINFEKL specific CD8+ T cells from KrasG12D+/-  Trp53-/- lung adenocarcinoma bearing lungs at different timepoints were sorted into 96-well plates with 5 µl lysis buffer comprised of Buffer TCL (Qiagen 1031576) plus 1% 2-mercaptoethanol (Sigma 63689). Following sorting, plates were spun down for one minute at 3,000 rpm and immediately frozen at −80°C. 10X: 10X Chromium Single Cell V(D)J kits (5' gene expression with paired T cell receptor sequencing, and for some samples Feature Barcode detection); Smart-Seq2: For preparation of single-cell libraries we thawed the cells and purified them with 2.2x RNAClean SPRI beads (Beckman Coulter Genomics) without final elution (Shalek et al., 2013). The RNA captured beads were air-dried and processed immediately for cDNA synthesis. We performed SMART-seq2 following the published protocol (Picelli et al., 2013).