Cfx connect real time pcr software

Total RNA was extracted from QF muscle and C2C12 cells using RNAiso Plus Total RNA Extraction Reagent (TaKaRa, Shiga, Japan), and cDNA was synthesized using the BioFact RT Series Kit (Biofact, Daejeon, Korea). Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) was performed to analyze gene expression with continuous monitoring using Bio-Rad CFX Connect Real-Time PCR software, version 1.4.1 (Bio-Rad Laboratories, Hercules, CA, USA). The gene-specific primer set (MyHC1, Tnni1, Tnnc1, MyHC2b, MyHC2x, and GAPDH) and protocols were from a prior study [40 (link)]. Gene expression levels were normalized to that of GAPDH.

Total RNA was extracted from RAW264.7 cells using RNAiso Plus Total RNA Extraction Reagent (TaKaRa, Shiga, Japan), and cDNA was synthesized using the BioFact RT Series Kit. Quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) was performed to analyze the expression of pro-inflammatory cytokines with continuous monitoring using Bio-Rad CFX Connect Real-Time PCR software, version 1.4.1 (Bio-Rad Laboratories, Hercules, CA, USA). The following primers were used: IL-1β forward, 5′-GAAGCGCTGCTTCCAAACCT-3′, IL-1β reverse, 5′-TGATGTGCTGCTGCGAGATT-3′, TNF-α forward, 5′-ACCGTCAGCCGATTTGCTAT-3′, TNF-α reverse, 5′-CTGGAAGACTCCTCCCAGGT-3′, IL-6 forward, 5′-TACCACTTC- ACAAGTCGGAGGC-3′, IL-6 reverse, 5′-CTGCAAGTGCATCATCATCGTTGTTC-3′. Expression levels were normalized to those of GAPDH.

HUVEC were treated with different compounds for 6h. Then total RNA was extracted from HUVEC using RNAeasy kit (beyotime, Shanghai, China) and cDNA was synthesized using the RT Master Mix for qPCR kit (MCE, New Jersey, USA). PCR amplification with a reaction volume of 10 μL, including 0.4 μL forward primer (10 μM), 0.4 μL reverse primer (10 μM), 5 μL SYBR, 3.2 μL RNase-free water and 1 μL cDNA was monitored by BIORAD CFX Connect Real-Time PCR Software, version 1.4.1 (Bio-Rad Laboratories, Hercules, CA, USA). The following primers were used: