Total RNA-seq: Sample H1_L45_DIV4
DRG were extracted from donors less than 2 hrs after aortic cross clamp, placed in NMDG solution for transport to culturing lab for fine dissection and minced. Pieces were dissociated enzymatically and mechanically and final solutions were filtered and cultured with DRG media (Valtcheva et al 2016). After 4 days in vitro, cultured coverslips were scraped into RNAlater, stored at -80 C and shipped on dry ice to sequencing center.Tissue was lysed using QIAzol, RNA extracted using RNEasy Plus Universal , and subjected to Illumina ribosomal RNA depletion and total RNA Tru-seq Gold library preparation protocol. Single end sequencing was done on the Illumina NextSeq platform.