GSM9761498: 2.4; Mus musculus; RNA-Seq
Mice were transcardially perfused on days 2 and 8 following chemotherapy or saline treatment using RNAlater (Invitrogen). Lumbar dorsal root ganglia (L3–L5, both sides) were dissected and total RNA was extracted using the RNeasy Mini Kit (Qiagen) with on-column DNase digestion according to the manufacturer's instructions. RNA-seq libraries were prepared using the Illumina Stranded Total RNA Prep Ligation with Ribo-Zero Plus kit and Illumina RNA UD Indexes according to the manufacturer's instructions. Briefly, 60–80 ng of total RNA underwent rRNA depletion and heat fragmentation prior to cDNA synthesis using random primers. Strand specificity was maintained using dUTP incorporation during second-strand synthesis. Libraries were ligated with indexed adapters, amplified by PCR, quantified using the Revvity LabChip GX Touch system with DNA High Sensitivity reagents, and pooled in equimolar ratios prior to sequencing.