2.6. Total RNA Extraction and Quantitative Real-Time PCR

The Total RNA was extracted from muscle samples using TRIzol reagent (Thermo Fisher Scientific Inc., Waltham, MA, USA) and the qualified RNA samples, which were determined by a NanoDrop 2000 (Thermo Fisher) instrument, were used for subsequent reverse transcription. The cDNA was synthesized by reverse transcription from RNA using random primers and a ReverAid First Strand cDNA Synthesis Kit (Thermo Fisher). The protocols and specific primers used for qPCR were mentioned, as previously described [12].