GSM6793119: MLM-TIL; Mus musculus; RNA-Seq
For analyses of infiltrative immune cells, single-cell suspensions (also see Flow cytometry) from APTKA PT, LM and PC samples (cells from 6 mice per location were pooled before) were subjected to single-cell sorting using a MoFlow Astrios machine. After exclusion of CD45- cells, all CD45+ / CD11B- cells were sorted into Eppendorf tubes and directly processed for 10X protocols. Single cell capture, reverse transcription, and library preparation were carried out on the Chromium platform (10X Genomics) with the single-cell 3' reagent v2 protocol according to the manufacturer's recommendations using 50,000 cells as input per reaction well. The final libraries [PT, LM, PC] were pooled and sequenced on two Illumina NovaSeq SP lanes (paired-end 26 bp + 96 bp).