title: GSM8114551: GBM mouse treated with oIL2 - rep1; Mus musculus; RNA-Seq
library_construction_protocol: After tumor dissociation, sample processing and staining were performed at 4°C. Cells were stained with anti-mouse FcγIII/II receptor (CD16/CD32) blocking antibody (1:100 dilution) for 10 minutes and then incubated in the presence of ⍺CD45.2-PB and ⍺CD45.1-APC-Cy7 antibodies for 30 minutes in the dark. 7-AAD was added to each sample just before sorting to label dead cells. CD45.2+ adoptive and both CD45.1+ and CD45.1+/CD45.2+ endogenous immune cells were then sorted using a BD FACSAria Fusion Cell Sorter (BD Biosciences), resuspended in 1% BSA-PBS at a concentration of 1000 cells/µL. Library was constructed using a microfluidics-based approach on Chromium Controller (10X Genomics) using the Chromium Next-GEM Single Cell 3' Reagent Kit v3.1 (10X Genomics) according to the manufacturer's instructions. Briefly, 16500 cells were added to each channel to achieve a recovery rate of 10000 cells per sample. Cells were partitioned in Gel Beads in Emulsion (GEMs, 10X Genomics) and lysed, followed by RNA barcoding, reverse transcription and PCR amplification (12 cycles). The concentration and size distribution of the scRNAseq libraries were determined using 4200 TapeStation system (Agilent).