GSM8061303: B16_PBS; Mus musculus; RNA-Seq
B16F10 tumor were harvested from tumor bearing mice as denoted and single-cell suspensions were preparation, viable CD45+ tumor infiltrating leukocytes were sorted by FACS Library was performed according to the manufacter's instructions (single cell3' v2 protocol, 10x Genomics). Briefly, GCs 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 llumina R1 primer sequence, Unique Molecular Identifier (UMI) and the 10x Barcode. The pooled barcoded cDNA was then cleaned up with Silane DynaBeads, amplified by PCR and the apropiated sized fragments were selected with SPRIselect reagent for subsequent library construction. During the library construction llumina R2 primer sequence, paired-end constructs with P5 and P7 sequences and a sample index were added.