GSM5385649: 2273_RNA; Homo sapiens; RNA-Seq
After FACS sorting and before encapsulation, cells were manually counted with hemacytometer and viability was also checked using Trypan blue exclusion. For each sample, 1,000-10,000 cells were loaded into the Chromium machine with the aimed recovery of 600-8,000 cells. Single cells were encapsulated into droplets with reagents and gel bead containing a unique molecular identifier (UMI) from 10x. cDNAs obtained after droplets break were purified with Dynabeads MyOne SILANE and amplified by 16 cycles of PCR (98°C for 45 s; [98°C for 20 s, 67°C for 30 s, 72°C for 1min] x 16; 72°C for 1min). Resulting amplified cDNAs were used for both 5'Gene Expression Library construction and V(D)J enriched libraries (TCR enrichment). Quantification of the resulting libraries was performed with the Qubit HS dsDNA assay kit and quality control was performed with the Fragment Analyzer (Agilent).