Rqg sybr green supermix

RNA was isolated from finely crushed snap‐frozen mouse quadriceps and heart samples using Trizol Reagent (Life Technologies), treated with DNAse (Promega; Madison, WI), and reverse transcribed using the SuperScript III kit (Life Technologies). Resulting cDNA was subjected to real‐time PCR using RQG SYBR Green supermix (Qiagen) in a Rotor Gene Q real‐time PCR machine (Qiagen). The following mouse‐specific primers were used: Fbxo32 (forward) 5′‐GCA GCC GCT CAG CAT TCC CA‐3′ and (reverse) 5′‐ACC GAC GGA CGG GAC GGA TT‐3′; Trim63 (forward) 5′‐AGG GCT CCC CAC CAC TGT GT‐3′ and (reverse) 5′‐TTG CCC CTC TCT AGG CCA CCG‐3′; Fbxo30 (forward) 5′‐ATC GAT GGC CCG TTA GTT ATT CA‐3′ and (reverse) 5′‐GCC CCT ATC TCA CCC TCA TCA AG‐3′; Acvr1b (forward) 5′‐GAA CCG CTA CAC AGT GAC CA‐3′ and (reverse) 5′‐ AAT TCC CGG CTT CCC TTG AG‐3′; Tgfbr1 (forward) 5′‐GCA TTG GCA AAG GTC GGT TT‐3′ and (reverse) 5′‐TGC CTC TCG GAA CCA TGA AC‐3′; Tgfb1 (forward) 5′‐GAC TCT CCA CCT GCA AGA CCA T‐3′ and (reverse) 5′‐GGG ACT GGC GAG CCT TAG TTT‐3′; Gapdh (forward) 5′‐AGC AGG CAT CTG AGG GCC CA‐3′ and (reverse) 5′‐ TGT TGG GGG CCG AGT TGG GA‐3′. Relative gene expression quantification was performed using the ΔΔC_t method with either Gapdh or Acvr1b as the reference gene.

RNA was isolated from finely crushed snap-frozen mouse quadriceps samples using Trizol Reagent (Life Technologies), treated with DNAse (Promega; Madison, WI), and reverse transcribed using the SuperScript III kit (Life Technologies). Resulting cDNA was subjected to real-time PCR using RQG SYBR Green supermix (Qiagen) in a Rotor Gene Q real-time PCR machine (Qiagen) with the following mouse-specific primers: Mstn (forward) 5’- GAC CCG TCA AGA CTC CTA CAA CAG-3’ and (reverse) 5’- ATC GCA GTC AAG CCC AAA GTC TCT-3’; Gdf11 (forward) 5’- GGC CGG CGT CAC ATC CGT ATC-3’ and (reverse) 5’- GCC GGA GCA GTA GTT GGC CT-3’; Inhba (forward) 5’- CAG GAG GGC CGA AAT GAA TGA ACT-3’ and (reverse) 5’- CCG CAC GTC CAG GGA ACT CTT T-3’; Cdkn1a (forward) 5’- GCT CAT GGC GGG CTG TC-3’ and (reverse) 5’- CTG CGC TTG GAG TGA TAG AAA T-3’. Primers for Gapdh [(forward) 5’-AGC AGG CAT CTG AGG GCC CA-3’ and (reverse) 5’-TGT TGG GGG CCG AGT TGG GA-3’] were used for ΔΔCt normalization.