Ssoadv univer sybr smx

Total RNA was isolated from the left ventricle using Total RNA Mini Plus (A&A Biotechnology, Gdynia, Poland) according to the manufacturer’s instructions. DNase-treated RNA (A&A Biotechnology, Gdynia, Poland) was reverse-transcribed using the RevertAid H Minus First Stand cDNA Synthesis Kit (Thermo Scientific, Pittsburgh, PA, USA). Real-time quantitative PCR was performed using the CFX Connect Real-Time PCR Detection System (Bio-Rad, Hercules, CA, USA). SsoAdv Univer SYBR SMX (Bio-Rad, Hercules, CA, USA) was used to detect and quantify mRNA expression. The relative expression of each sample was determined after normalization to β-actin or 60S ribosomal protein L32 (RPL32) using the ΔΔCt method. A list of primers for real-time PCR is presented in Table 2.

Total RNA was isolated from the LV using Total RNA Mini Plus (A&A Biotechnology, Gdynia, Poland) according to the manufacturer’s instructions. DNase-treated RNA (A&A Biotechnology, Gdynia, Poland) was reverse-transcribed using the RevertAid H Minus First Stand cDNA Synthesis Kit (Thermo Scientific, Pittsburgh, PA, USA). Real-time quantitative polymerase chain reaction (PCR) was performed using the CFX Connect Real-Time PCR Detection System (Bio-Rad, Hercules, CA, USA). SsoAdv Univer SYBR SMX (Bio-Rad, Hercules, CA, USA) was used to detect and quantify mRNA expression. The relative expression of each sample was determined after normalization to β-actin or 60S ribosomal protein L32 (Rpl32) using the ΔΔCt method. A list of primers for real-time PCR is presented in Supplementary Table S1.