title: GSM8504103: IPD5,male, 50d; Homo sapiens; RNA-Seq
library_construction_protocol: Midbrain organoids were collected from their culture medium and washed with 1x PBS (phosphate-buffered saline, Gibco, Cat#10010-015). Organoids were transferred to a 1.5ml Eppendorf tube and digested in 1 ml sCelLive™ Tissue Dissociation Solution (Singleron Biotechnologies, Cat#1190062) diluted 1:2 with PBS. The organoids were placed in a thermal shaker at 750 rpm at 37°C for 45 minutes. The state of dissociation was checked at regular intervals under a light microscope. Following digestion, the suspension was filtered using a 40-µm sterile strainer (Greiner, Cat#542040). The cells were centrifuged at 350xg for 5 minutes at 4°C, and the cell pellets were resuspended in 300µl PBS. Cells were stained with Acridine Orange/Propidium Iodide Stain (Logos Biosystems, Cat#F23001), and the cell number and viability were calculated using LUNA-FX7™ Automated Cell Counter (Logos Biosystems, Villeneuve d'Ascq, France). The single-cell RNA-seq libraries were constructed using GEXSCOPE™ Single Cell RNAseq Library Kit (Singleron Biotechnologies, Cat#4180011) according to the manufacturer´s instructions.