GSM8866060: T2Day1; Homo sapiens; RNA-Seq
Endometrial organoids were harvested using cell recovery solution, incubated on ice for 30 minutes, and centrifuged at 400 ×g for 3 minutes to remove supernatant. The organoids were digested with a mixture of 10% DNase and TrypLE Express at 37°C for 5 minutes, followed by termination with 10% FBS. After filtering the cell suspension through a 35 µm strainer (Falcon, 352235) to remove aggregates, dead cells were selectively removed using Dead Cell Removal Beads (Miltenyi, 130-090-101) and MACS separation with a LS Column and MACS Separator (Miltenyi, 130-090-312). Viable cells were resuspended in 4°C PBS. Samples meeting viability (>80%) and density criteria (1000-5000 cells, 500 cells/μl at 0.5×10⁶ cells/ml) were submitted for sequencing analysis at the Centre for PanorOmic Sciences, HKU. Library preparation and Illumina sequencing (Pair-End sequencing of 150 bp) were conducted at the Genomics Core, Centre for PanorOmic Sciences (CPOS), LKS Faculty of Medicine, the University of Hong Kong. Cells will be encapsulated into Gel Beads-in-emulsion (GEMs) by the 10X Chromium iX, followed by reverse transcription and library preparation to become a pool of cDNA libraries. Transcripts from the same cell will be identified by a 10X barcode, while each of these transcripts will be discernible from one another by a Unique Molecular Identifier (UMI).
GEO
GSM8866060_r1