GSM8353974: DRG: Naive, 0 days rep 1; Rattus norvegicus; RNA-Seq
DRG were cryosectioned at 20µm thickness onto polyethylene-naphtalate (PEN) membrane slides. Sections were briefly dehydrated and then laser dissected using a PALM MicroBeam (Carl Zeiss, Germany) and PALM Robosoftware. Large diameter (>40µm) neurons labelled with fluorophore/ tracer were dissected and catapulted into an adhesive cap. An average of 184 neurons per animal were collected. Trizol Reagent was added to the samples. To isolate RNA, chloroform extraction was performed and then the RNeasy® Micro Kit (QIAGEN, Germany) was used following an adapted version of the manufacturer's protocol. RNA quality numbers, determined by Fragment Analyzer, were in the range 5.5-8.1. The NEBNext Ultra Directional RNA Library Prep Kit for Illumina was used, following the manufacturer's protocol (NEB #E7420S/L), in conjunction with ribosomal RNA depletion.