GSM8989595: D1 sorted cells, Saline-Saline, Replicate 2; Mus musculus; RNA-Seq
Nuclei were purified and nuclei were then immediately sorted using a BD FACS Aria II three-laser system with a 100um nozzle. Debris and doublets were first gated based on forward scatter and side scatter, nuclei were then gated as DAPI-positive events (Violet1-A laser), and GFP positive events were sorted directly into 1.5 ml Eppendorf tubes containing TriZol LS (Ambion) and flash frozen on dry ice. Samples ranged from 18,000-60,000 GFP+ nuclei collected for downstream analysis. Processed with Chromium Next GEM Single Cell 3′ GEM, Library & Gel Bead Kit v3 according to the manufacturer's instructions to construct the scRNA-seq library.