GSM8267315: Control_western_diet_rep2; Mus musculus; RNA-Seq
RNA was extracted from skeletal muscle harvested from control and transgenic mice using TRI-reagent (Molecular Resource Center), followed by in-column DNase-digestion using RNeasy kit (Qiagen). Samples were processed according to the manufacturer's instructions. RNA concentration was determined using a NanoDrop spectrophotometer (Thermo Fisher Scientific), and RNA integrity using an Agilent BioAnalyzer 2100 (Agilent Technologies). Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers, followed by the second strand cDNA synthesis. The library was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. Quantified libraries will be pooled and sequenced on Illumina platforms, according to effective library concentration and data amount.