title: GSM7112547: DCP-IL-12/FLT3L replicate 2; Mus musculus; RNA-Seq
library_construction_protocol: Subcutaneous B16F10 tumors were enzymatically digested by tumor dissociation kit (Miltenyi Biotec; Cat. No. 130-096-730) in gentleMACS™ Octo Dissociator machine (Miltenyi Biotec; Cat No.130-096-427) Single-cell tumors were then washed twice in PBS containing 0.04% BSA. After filtration through a 40 um Flowmi strainer (Bel-Art, Wayne, NJ), cells were resuspended in PBS with 0.04% BSA, checked for absence of significant doublets or aggregates, and loaded into a Chromium Single Cell Controller (10x Genomics, Pleasanton, CA) in a chip together with beads, master mix reagents (containing RT enzyme and poly-dt RT primers) and oil to generate single-cell-containing droplets. Single-cell gene expression libraries were then prepared using Chromium Single Cell 3' Library & Gel Bead Kit v3.1 (PN-1000268) following the manufacturer's instruction (protocol CG000315 Rev C). Quality control was performed with a TapeStation 4200 (Agilent) and QuBit dsDNA high sensitivity assay (Thermo) following manufacturer instructions. The sequencing libraries were loaded onto an Illumina HiSeq 4000 paired-end Flow Cell and sequenced using read lengths of 28 nt for read1 and 91 nt for read2, at a depth of ca 35k reads/cell.