RNA-seq of mouse: glKO ACC 4hr Spinal Cord 1
RNA was extracted and purified from SC tissue samples using the Total RNA Purification Kit (Norgen Biotek Corp., Canada) following manufacturers instruction. RNA sequencing was performed by Novogene (California). Briefly, mRNA was separated out using poly-T oligo-attached magnetic beads. The strands were fragmented, then one round of reverse transcription was performed with random hexamer primers to synthesize cDNA. A second strand cDNA synthesis was performed using dUTP for directional library sequencing. End repair and A-tailing were performed followed by Qubit and real-time PCR for quantification. Adapters were ligated to each strand and a bioanalyzer completed the size distribution selection. Sequencing was finished on Illumina platforms.