GSM8030994
Fresh murine spleens were submitted to mechanic disaggregation with a syringe plunger through a 70 µm cell strainer, followed by lysis of erythrocytes with ACK buffer (NH4Cl 150 mM KHCO3 10 mM EDTA 0.1 mM). Cell suspensions were sorted in a FACSAria II sorter to obtain a final cell suspension of 20% live lymphoma 7AAD- GFP+ cells and 80% microenvironment 7AAD- GFP- cells. For each sample, 17,000-19,000 cells were loaded on a Chromium Single-Cell Instrument (10X Genomics) at a concentration of 721 cells/µL to generate single-cell gel bead-in-emulsions (GEMs) of approximately 10,000 cells per sample. The Chromium Next GEM Single Cell 5' Reagent Kit v2 (Dual Index) from 10X Genomics was used to produce libraries for both sequencing-ready Gene Expression and T cell V(D)J libraries.