GSM9571270: Female Down syndrome,organoid; Homo sapiens; RNA-Seq
Human fetal brain tissue (24 GW) or organoid (2.5 months culture) are minced and homogenized using a Dounce homogenizer on ice, then washed and filtered through a 40 μm mesh to generate a clean nuclear suspension ready for the 10x Genomics Chromium platform. snRNA seq libraries were prepared using the 10× Genomics Chromium Controller and Chromium Single Cell 3′ V3.1 reagent kit (10× Genomics, Pleasanton, USA). Isolated nuclei were adjusted to a concentration of 1000 nuclei per uL and loaded onto the Chromium chip to generate single-nucleus Gel Bead-In-Emulsions (GEMs). Reverse transcription was performed within GEMs, followed by GEM disruption and recovery of barcoded cDNA, which was then purified and amplified. The amplified cDNA was fragmented, a tailed, adaptor ligated, and further enriched by index PCR. Library concentration was determined using the Qubit High Sensitivity DNA assay (Thermo Fisher Scientific, USA), and fragment size distribution was assessed with a High Sensitivity DNA chip on a 2100 Bioanalyzer (Agilent, USA). All libraries were sequenced on a NovaSeq 6000 platform (Illumina, USA) using a 150 bp paired end configuration.