GSM7574586: KPT scRNA-Seq; Mus musculus; RNA-Seq
Tumours were dissected from mice on day 14 after inoculation and disassociated using the Miltenyi Mouse Tumour Dissociation Kit and gentleMACS Octo-Dissociator (Miltenyi) as per the manufacturer's instructions. Isolation of immune cells was performed by density centrifugation with Percoll. Cells were stained using anti-CD45 APC (BioLegend, 103112, 1:200), LIVE/DEAD FITC (Invitrogen, L34969, 1:1000) antibodies followed by selection of live CD45 positive single cells with FACS. Cell capture and cDNA synthesis was using single cell 3'libray and Gel Bead kit V3.1 (10x Genomics, 1000121) and Chromium single cell G Chip Kit (10x Genomics, 1000120) according to the manufacture's introduction. Single cell RNA-seq libraries were constructed using Single Cell 3' Library and Gel Bead Kit V3.1.