GSM9634916: Gel+US spinal cord lesion core biol rep 3; Rattus norvegicus; RNA-Seq
Spinal cord tissue (~5 mm3) from the lesion epicenter (T9 segment) was harvested. Total RNA was extracted using the TRIzol method, and RNA integrity was assessed using an Agilent 2100 Bioanalyzer. Poly(A)+ mRNA was enriched from total RNA using Oligo(dT) magnetic beads. The purified mRNA was fragmented in NEB Fragmentation Buffer and reverse transcribed using random hexamer primers to generate first-strand cDNA. Second-strand cDNA was synthesized using dUTP instead of dTTP to preserve strand information. After end repair, A-tailing, adaptor ligation, fragment size selection, USER enzyme digestion, PCR amplification, and purification, strand-specific libraries were generated for sequencing.