GSM7908792: hNMSO_d39-44_spatial; Homo sapiens; OTHER
For spatial transcriptome sequencing, organoids were embedded in OCT compound and sectioned at 10 μm thick and were placed on the capture area of the Visium Spatial Gene Expression Slide. For spatial transcriptome sequencing, Visium Spatial Gene Expression Reagent Kits were applied according to the manufacturer's instructions. Sections were fixed with formaldehyde, Hematoxylin and Eosin staining were performed, and sections were scanned for collection. Capture Area contains ~5000 gene expression spots that include primers including Illumina TruSeq Read 1, 16 nt Spatial Barcode, 12 nt UMI, and 30 nt poly(dT) sequence required for capture and priming of poly-adenylated mRNA. Once the samples were permeabilized, RT Master Mix containing reverse transcription reagents was added to the permeabilized sections. Cellular mRNA was captured by the primers on the gene expression spots to generate full-length cDNA. Libraries were generated with P5, P7, i7, and i5 sample index, and TruSeq Read 2 was added to each selected cDNA for sequencing.