title: GSM6542232: CD8 T Cells from MafBCreIRF8FLPyMT Tumor, scRNAseq; Mus musculus; RNA-Seq
library_construction_protocol: Around 10,000 FACS sorted live cells (1000 cells/ml suspension, more than 90% viability evaluated by DAPI nuclear stain) were suspended in 1X PBS (calcium and magnesium free) containing 0.04% BSA (A2153, SIGMA). Cell suspension was used as input in 10x chromium controller system (10x Genomics Inc., product code 120223) and cell barcoding was performed by gel beads in emulsion (GEMs) in assembly chip. GEM RT reaction was performed in thermocycler ( 530C for 45 minutes, 850C for 5 minutes, 40C hold overnight). SPRIselect dynabeads (B23318, Beckman Coulter) were used for GEM recovery post RT 2-50ng of DNA was used for target enrichment. cDNA amplification, fragmentation, end repair and A tailing preparation was performed as per manual instructions. High sensitivity DNA chips and Agilent 2100 bioanalyzer (Agilent Technologies) was used for gene expression library profile quality control and quantification at several steps. Quality control was performed twice before sequencing. Two barcoded scRNA samples were pooled together before sequencing