title: GSM7796986: Aged_CD8, scRNAseq; Mus musculus; RNA-Seq
library_construction_protocol: Tumor tissues were collected and cut into small pieces (approximately 1-2 mm). Then, we digested the tumor tissues with a mouse tumor dissociation kit (Miltenyi/MACS, 130-096-730) following the standard procedure. After that, the cell suspension was filtered with a 30 μm MACS SmarterStrainer. Then, we performed either FACS analysis or FACS sorting (sorting for CD45+CD8+T cells). The cell suspension samples we obtained with this procedure were used for single-cell library construction. We performed single-cell immune profiling following the standard procedure from 10X Genomics. .Briefly, CD8+T cells were resuspended in the master mix and loaded together with partitioning oil and gel beads into the chip to generate the gel bead-in-emulsion (GEM).The poly-A RNA from the cell lysate contained in every single GEM was retrotranscripted to cDNA, which contains an Ilumina R1 primer sequence, Unique Molecular Identifier (UMI) and the 10X Barcode.The pooled barcodeed cDNA was then cleaned up with Silane DynaBeads, amplified by PCR and the apropiated sized fragments were selected with SPRIselect reagent for the subsequent library construction. During the library construction Illumina R2 primer sequence, paired-end constructs with P5 and P7 sequences and a sample index were added.