Taq pro universal sybr qpcr master mix

Thyroids were gently dissociated and then stored at -80°C. Total ribonucleic acid (RNA) was extracted with TRIzol reagent. The concentrations of RNA were determined by measuring absorbance at 260 nm using a UV5 Nano spectrophotometer (Mettler Tolero, Switzerland). Complementary deoxyribonucleic acid (cDNA) was prepared with a HiScript III RT SuperMix reverse transcriptase kit by a T100 thermal cycler system (Bio-Rad, Hercules, CA, USA). Quantitative reverse transcription polymerase chain reaction was conducted with a Taq Pro universal SYBR qPCR master mix by an ABI 7500 real-time PCR system (Applied Biosystems, Waltham, MA, USA). Rat-specific PCR primers were designed. The sequences were as follows: Il-17a, forward 5’-CTC AAC CGT TCC ACT TCA CC-3’, reverse 5’-CAC TTC TCA GGC TCC CTC TTC-3’; Traf6, forward 5’-CCC AAG AAG AGG AAA GAC-3’, reverse 5’-AGG ATC GTG AGG CGT AT-3’; Erk1, forward 5’-CTG GCA CTG AAG GAG G-3’, reverse 5’-AAC AAG ATG AGG CTA CG; Erk2, forward 5’-CCA GAG TGG CTA TCA AGA AG-3’, reverse 5’-GGA TGT CTC GGA TGC CTA; Tnf-α, forward 5’-CCA CCA CGC TCT TCT GTC T-3’, reverse 5’-GCT ACG GGC TTG TCA CTC G-3’; Gapdh, forward 5’-CTC TGC TCC TCC CTG TTC-3’, reverse 5’-CGA TAC GGC CAA ATC C-3’. Gapdh was used as an endogenous control. The PCR results were expressed as the relative expression ratio of targeted genes and Gapdh.