title: NextSeq 500 paired end sequencing; Single-cell RNA-seq (10x Chromium V3) of heterogeneous spheres consisting of melanoma cell line and two types of human primary dermal fibroblasts
design: Single-cell RNA-seq (10x Chromium V3) of heterogeneous spheres consisting of melanoma cell line and two types of human primary dermal fibroblasts
library_construction_protocol: Heterogeneous spheres were prepared from primary human fibroblasts and G361 melanoma cell line (commercially available; CVCL_1220). Fibroblasts were isolated from photo-aged skin of an adult and from juvenile photo-protective skin of child. Heterogeneous spheres were washed in PBS and digested in 0.25% trypsin + EDTA solution (1:1). Cell suspensions were inoculated in DMEM + 10% fetal bovine serum for 72 hours in hanging drops. Heterogeneous spheres were cultured for additional 24 hours in non-adhesive culture dishes.  Immediately after tissue dissociation, single cell libraries were prepared on the 10x Chromium with a target of 4,000 cells. Chromium Single Cell 3สน GEM, Library & Gel Bead Kit v3, 4 rxns PN-1000092 was used. Immediately after tissue dissociation, single cell libraries were prepared on the 10x Chromium with a target of 4,000 cells. Chromium Single Cell 3สน GEM, Library & Gel Bead Kit v3, 4 rxns PN-1000092 was used.