GSM6732801: Lymphoid compartment - treated with anti-CD25 - Second replicate; Mus musculus; RNA-Seq
Tumors were microdisected from the brain to avodi contamination with normal tissue and subsequently enzymatically digested to obtain a single cell suspension ready to perform single cell sorting Single-cell suspensions were prepared and Live CD45-, Live CD45+CD3+ and Live CD45+CD11b+ populations were sorted. Cells were washed, counted and resuspended to a concentration of 1000 cells/uL in PBS 0.04% BSA. After this, cells were loaded and processed on the 10X Chromium Controller using the CG000331 Chromium Next GEM Single Cell 5' Reagent Kits v2 (Dual Index) User guide, which enabled us to process the gene expression and immune profiling of the samples. Once the libraries were processed, they were pooled with a GEX 4:1 TCR ratio (as per 10X recommendation) and submitted for sequencing. Each library can be recognized by its unique sample index. Sample sequencing was performed at Novogene, using a Novaseq 6000 platform and PE150 sequencing strategy.