GSM3589963: MOO2450A24: Rat 61; Rattus norvegicus; RNA-Seq
Total RNA for RNA sequencing was extracted from C6-C8 DRGs on the affected side for each rat. Tissue was homogenized in QIAZOL (Qiagen, 79306) and nucleic acids were separated in Phase Locked Gel columns (5Prime, 230 2830). 1.5 volume of 100% ethanol was added to the aqueous phase and transferred to filtered spin columns from the RNA extraction kit (miRNeasy kit, Qiagen, 217004) and we proceeded according to manufacturer's instructions. Samples were also DNase I treated with double volume of the recommended amount (Qiagen, 79254). We estimated the total RNA quality and quantity by spectrophotometry (NanoDrop, ND-1000) and measured the RIN scores (Agilent RNA 6000 Nano Reagents Part I and Agilent 2100 Bioanalyzer). The integrity number for each sample was greater than 7.6, with an average of 8. The samples were subjected to poly-A enrichment with oligo-dT beads (Illumina) and the library was prepared using the TruSeq Stranded prep kit (Illumina)