Abi stratagene mx3000p instrument

The total RNA was extracted from the skin of the KO and control groups using TRIzol reagent (Thermo Fisher Scientific, ShangHai). PrimeScrip RT reagent Kit with gDNA Eraser (Perfect Real Time) (Takara Biomedical Technology, Beijing) was used to obtain the cDNA. Real-time PCR was performed on ABI Stratagene Mx3000P instrument (Agilent Technologies, Santa Clara, CA, USA) using a TB Green Premix Ex Taq II (Takara Biomedical Technology, Beijing). The primer sequences used in this experiment are shown in Supplementary Table S3. The expression levels were analyzed using the 2^ΔΔCt method, and normalized against GAPDH. Each sample was run in triplicate.

Total RNA was extracted from gluteal muscle tissue of sheep from MSTN-knockout (#44 and #48) and control individuals (#06, #24, and #38) using TRIzol™ reagent (Thermo Fisher Scientific, Shanghai, China). PrimeScrip™ RT Reagent Kit with gDNA Eraser (Perfect Real Time; Takara Biomedical Technology, Beijing, China) was used to obtain cDNA. Real-time PCR was performed in ABI Stratagene Mx3000P instrument (Agilent Technologies, Santa Clara, CA, USA) using TB Green Premix Ex Taq II (Takara Biomedical Technology). Primer sequences used in this experiment are listed in Additional file 1: Table S8. Gene expression levels were calculated using the 2^-ΔΔCt method and normalized against housekeeping GAPDH gene. Each sample was run in triplicate.