Instrument: NextSeq 500
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Sciatic nerves and the brachial nerve plexus were dissected from intracardial PBS perfused the mice. Nerves were first mechanically dissociated. Cells were then isolated by combining enzymatic digestion (Trypsin, Collagenase II, Hyaluronidase and Pronase) with myelin depletion and flow cytometry-based cell sorting (Calcein-AM, Zombi NIR, DAPI). Rats sciatic nerves were isolated and enzymatically digested in Collagenase/Dispase and DNase I. Cells were centrifuged on a 30%/70% Percoll gradient and the mononuclear interphase was collected. No sorting was performed. Single cell suspensions were loaded onto the Chromium Single Cell Controller using the Chromium Single Cell 3' Library & Gel Bead Kit v2 (both from 10X Genomics) chemistry following the manufacturer's instructions. Sample processing and library preparation was performed according to manufacturer instructions using AMPure XP beads (Beckman Coulter). Sequencing was either carried out on a local Illumina Nextseq 500 using the High-Out 75 cycle kit with a 26-8-0-57 read setup or commercially (Microanaly, China) on a NovaSeq 6000 using the 300 cycle kit with paired end 150 read setup. All the samples were sequenced with a sequencing depth >50,000 reads per cell ScRNA-seq