GSM9450696: QBM-23; Mus musculus; RNA-Seq
Uveal melanomas were harvested from GnaqCA; Bap1CKO; MycCKI mice when ocular enlargement was visibly apparent. Mice were euthanized, and eyes were enucleated using fine scissors and forceps under a dissection microscope. After removal, each eye was placed in ice-cold sterile PBS. Under a stereomicroscope, extraocular tissues (conjunctiva, optic nerve), lens, and vitreous were removed. Tumor tissue arising from the choroid and adjacent structures was dissected out with fine-tipped forceps and immediately transferred into the enzyme solution. Enzymatic dissociation was performed using the Tumor Dissociation Kit (Miltenyi Biotec, RRID:SCR_020285) according to the manufacturer's instructions. Tissues were incubated at 37°C for 30–45 minutes with intermittent gentle agitation. The resulting cell suspension was washed with PBS containing 2% FBS and centrifuged at 300 x g for 5 minutes. Cells were resuspended in sterile PBS, Cell viability and counts were obtained by AO/PI dual fluorescent staining and visualization on the Nexcelom Cellometer K2 (Nexcelom Bioscience LLC, RRID:SCR_018656). Cell viability >80% was used for inclusion in sequencing. 10x genomics Gel Bead-in-Emulsion (GEM) library construction protocol