GSM5703304: Organoid_Line2_Dex_rep2; Homo sapiens; RNA-Seq
Single cells were dissociated using StemPro Accutase Cell Dissociation Reagent (Life Technologies), filtered through 30uM and 20uM filters (Miltenyi Biotec) and cleaned of debris using a Percoll (Sigma, P1644) gradient. Single cells were resuspended in ice-cold Phosphate-Buffered Saline (PBS) supplemented with 0.04% Bovine Serum Albumin and prepared for single-cell separation. All experiments were performed in quadruplicate using a pair-wise design with a treatment (Dex) or control (Veh) condition ran in parallel. Single cells were run through the Chromium controller to form gel emulsion beads containing barcoded single cells and prepared into single-cell libraries using the Chromium Single Cell 3' Reagent Kits v2 according to the manufacturer's recommendations without any modifications (10X Genomics). All libraries were assessed using a High Sensitivity DNA Analysis Kit for the 2100 Bioanalyzer (Agilent) and KAPA Library Quantification kit for Illumina (KAPA Biosystems). Sequencing of the 10x Genomics single-cell libraries was performed on an Illumina NovaSeq (Illumina, San Diego, CA), generating paired-end reads of 100 bp length.