GSM5133479: Naïve LN2; Mus musculus; RNA-Seq
The primary tumors were cut into 1-2 mm pieces and dissociated into single cells. All analyzed lymph nodes were dissociated into single cells with digestion buffer (200 ug/ml collagenase I, 800 ug/mL Dispase, 100ug / ml DNase I). All samples used ACK buffer (Thermo Fisher) to lyse red blood cells and had the cell concentration and viability tested using a Nadia instrument (Dolomite Bio). The loading quantity of viable cells for 10x Genomics platform was estimated to capture around 5000 cells. 10x genomics Chromium Single Cell 3' Reagent (v2 Chemistry) was used for library preparation. 10x genomics single-cell mRNA sequencing.