GSM7592783: Interferon gamma CMO302; Mus musculus; RNA-Seq
Spheroids were dissociated in 2 ml digestion medium (RPMI medium (Gibco) + 25 µg/ml Liberase ™ (Roche) + 40 µg/ml DNase I (Roche; 2000 U/ml) and incubated for 30-45 min in a water bath at 37°C with gentle shaking every 5 min. 4 ml PBS + 0.04% BSA was added to stop the digestion. Cell Multiplexing Oligos (CMO) labels B301 - B305 (3' CellPlex Kit Set A) were used to label the 5 samples. After labeling, cell suspensions were counted again and the 5 samples were pooled according to the pooling calculations in appendix of the labeling protocol (CG00391) in equivalent ratios. The pool was then directly used for GEM generation following 10X Genomics protocol (CG000388 Demonstrated Protocol Chromium Next GEM Single Cell 3ʹ v3.1 (Dual Index) with Feature Barcode technology for Cell Multiplexing Rev B) targeting a cell recovery rate of 10'000 cells.