title: Transcriptomic profiling of BrO, DMGO and GD2 CAR T cells
design: Transcriptomic profiling of BrO, DMGO and GD2 CAR T cells
library_construction_protocol: We performed single cell dissociation using the Neural Tissue Dissociation Kit (P) (Miltenyi Biotec, Cat. #130-092-628), adjusted for DMGOs. After cutting the organoids into smaller pieces and adding the enzyme mixes, the samples were incubated on an orbital shaker at 37°C and resuspended in regular intervals with a P1000 until a single cell suspension was reached, which was verified under the microscope using trypan blue to check for cell death. After dissociation, the single cell suspension was washed twice with PBS +/+ (magnesium/calcium+ 3% FBS). Next, DAPI was used as a viability dye (DAPI 1:5000). Stained single cells suspensions were filtered using a 40 µm Flowmi cell strainer and sorted by FACS on DAPI exclusion and enriched for GFP expression for tumour cells.  Nuclei acid extraction was provided by Leiden Genome Technology Center (Leiden, The Netherlands) Library generation was provided by Leiden Genome Technology Center (Leiden, The Netherlands)