GSM5752842: CD8+PD-L1-, rep2; Homo sapiens; RNA-Seq
After the stimulation, cells were surface stained for CD8, and PD-L1, and CD8+PD-L1+ and CD8+PD-L1- T cells were sorted using a BD FACSAria II cell sorter. The purity of the sorted samples was greater than 95%. Cells were sorted into tubes containing lysis buffer, and and total RNA were extracted by using the Quick-RNA Microprep Kit (Zymo). Then transcriptome amplifications were performed by using Single Cell Full Length mRNA-Amplification Kit (Vazyme). The amplified cDNA products were purified with DNA beads (KAPA) to eliminate short fragments (less than 500 bp). At this step, the concentration of each sample was quantified with Qubit HsDNA kits (Invitrogen), and libraries were then constructed with the TruePrep DNA Library Prep Kit V2 for Illumina (Vazyme). Finally, the libraries were sequenced on an Illumina Nova6000 sequencer (Illumina).