design: scRNA-seq time course of cerebellar Nestin-expressing progenitors from neonatal mice with and without injury at birth
library_construction_protocol: The single-cell RNA-Seq of FACS-sorted cell suspensions was performed on Chromium instrument (10X genomics) following the user guide manual for 3′ v3.1. In brief, FACS-sorted cells from different conditions were multiplexed using CellPlex reagents (10x Genomics) as described by the manufacturers protocol. The viability of cells was above 80%, as confirmed with 0.2% (w/v) Trypan Blue staining and barcoded cells were pooled into a single sample in PBS containing 1% BSA to a final concentration of 700–1,300 cells per μl. Between 2,000 to 3,000 cells were targeted for each sample. Samples were multiplexed together on one lane of 10X Chromium following 10x Genomics 3' CellPlex Multiplexing protocol and a total of ~30,000 cells were targeted for droplet formation. Following reverse transcription and cell barcoding in droplets, emulsions were broken, and cDNA purified using Dynabeads MyOne SILANE followed by PCR amplification per manufacturer's instruction.