title: GSM5172673: DJC10 Spleen-Mouse02; Mus musculus; RNA-Seq
library_construction_protocol: Samples were pre-enriched for total T cells by immunomagnetic negative selection using the StemCell Technologies EasySep Mouse T Cell Isolation Kit (19851) following the manufacturer protocols for all three peripheral tissue suspensions, with minor modifications for processing of tumor suspensions. The negative selection antibody cocktail for tumor samples were supplemented with 15ug/ml anti-CD34-biotin (eBioscience, 13-0341-82) to deplete MC38 tumor cells. After labeling, the tumor-cocktail suspension was washed once with 5 mL MACS buffer prior to a 10-minute incubation with eBioscience MagniSort Streptavidin Negative Selection Beads (Invitrogen, MSNB600274) at 20 μL beads/100 mL sample. All sample volumes were brought up to 2.5 mL with additional MACS Buffer. Samples were then placed on a StemCell Technologies magnet for 10 minutes, and T cells were subsequently collected. All enriched mouse single T cell samples were stained for viability (Sytox Red) and with antibodies against CD45, TCRβ, and CD90.2 and sorted via Aria sorters prior to loading onto the 10X genomics single cell platform according to the manufacturer protocol. Libraries were prepared using the 10X Chromium Single Cell 3' v3 Gene Expression Kit.