GSM5060422: CD45+ tumorinfiltrating immune cells_NL-_and_L-PM; Mus musculus; RNA-Seq
At day 15 single cell suspensions were prepared from excised whole tumors by tissue digestion using murine tumor dissociation kit and gentleMACS dissociator (Miltenyi Biotec). CD45+ cells were enriched by murine CD45 (TIL) MicroBeads (Miltenyi Biotec) using MACS technology. To enrich purity viable CD45+ cells were sorted using AriaIII flow cytometer (BD Bioscience). Cells derived from 3 independent experiments per group (3x NL-PM and 3x L-PM) were "labeled" by Sample Tags 3,4,5 (NL-PM) and 6,7,9 (L-PM) and then together loaded on the Rhapsody cartridge. After capturing of single cells RNA molecules bound to magnetic beads, libraries were generated following the BD Rhapsody System mRNA WTA and Sample Tag Library Preparation protocol (BD Biosciences). Barcoding of RPE and Sample Tag library was done by Library Reverse Primer 1 (Illumina i7 Index): 5'-GCTACGCT-3'