title: single-cell RNA sequencing of iAstrocytes from iPSC1: Day21
design: Astrocytes were generated from multiple iPSCs (iPSC1 and iPSC2) by overexpressing NFIB and SOX9. The resulting astrocytes derived from iPSC1(DYR0100) and iPSC2 (BIONi037-A) were profiled by scRNA-seq. Moreover, the resulting cells at multiple stages (Day 0, Day 1, Day 3, Day 8, Day 14, Day 21) along monocolonal iPSC1 differentiation were profiled by scRNA-seq. The Chromium Next GEM Single Cell 3 Reagent Kit v3.1 & Gel Beads Kit (10x Genomics) were used according to the manufacturers guidelines. Briefly, single-cell suspensions, gel beads and partitioning oil were loaded onto the 10x Genomic Chromium Chip (Next GEM chip G) and subjected to the 10x Chromium Controller device during which single cells were encapsulated with individual Gel Beads-in-emulsion (GEMs). The number of cells targeted in each experiment was 18000. Captured cells were lysed and the transcripts inside the individual GEMs were barcoded through reverse transcription. Constructed 10x libraries were quantified using a Qubit 4 Fluorometer (Invitrogen) with a Qubit 1x dsDNA HS Assay Kit (Invitrogen). Quality control for the 10x libraries was performed on a Fragment Analyzer 5200 (Agilent) with a DNF-474 HS NGS Fragment Kit (Agilent).