GSM8606551: 7KQ, replicate2,scRNAseq; Mus musculus; RNA-Seq
1×106 MC38 cells were inoculated into the flanks of the indicated mice and allowed to grow for approximately 19 days. Tumors were isolated and digested with 1 mg/ml collagenase I (Merck, C0130) and 200 μg/ml DNase I (Sigma, DN25), and the live Cd45+ tumor-infiltrating cells were sorted via flow cytometry. Library was performed according to the manufacter's instructions (Single Cell 3' Reagent Kits v3.1 (Dual Index, 10× genomics, 1000268). Briefly,  Cd45+ cells were resuspended in the master mix  and loaded together with partitioning oil and gel beads into 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 IIumina R1 pimer sequence, Unique Molecular Identifier (UMI) and 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 IIumina R2 primer sequence, paired-end constructs with P5 and P7 sequences and a sample index were added.