cDNA was generated from 1–5 µg of total RNA using High Capacity RNA-to-cDNA kit (Life Technology Grand Island, NY) according to the manufacturer’s instruction. qRT-PCR assays were performed using PerfeCT Super Green Super Mix (Quanta Biosciences, Beverly, MA) by using specific primers (Invitrogen, Carlsbad, CA) Differences in expression were determined by the relative quantification method; the cycle threshold (CT) values of the target genes were first normalized to the CT values of endogenous control GAPDH.
Perfect super green super mix
Manufactured by Quanta Biosciences
The PerfeCT Super Green Super Mix is a laboratory reagent designed for qPCR (quantitative Polymerase Chain Reaction) experiments. It contains all the necessary components, including a proprietary green fluorescent dye, for efficient and sensitive real-time PCR amplification and detection. The product is formulated to provide reliable and consistent results in gene expression analysis and other qPCR applications.
Automatically generated - may contain errors
6 protocols using perfect super green super mix
1
cDNA Synthesis and qRT-PCR Analysis
2
Quantifying Cardiac Metabolic Gene Expression
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Total RNA were extracted from LV tissue with TRIZOL. cDNA was generated from 2μg of total RNA using High Capacity RNA-to-cDNA kit (Life Technology Grand Island, NY) according to the manufacturer’s instruction. qRT-PCR assays were performed using PerfeCT Super Green Super Mix (Quanta Biosciences, Beverly, MA) by using specific primers PPAR-γ, HMGCS2, BDH1 and PDK4 (Invitrogen, Carlsbad, CA) Differences in expression were determined by the relative quantification method; the cycle threshold (CT) values of the target genes were first normalized to the CT values of endogenous control GAPDH. All primer sequences are given in Table 1 .
3
RNA-to-cDNA Conversion and qRT-PCR Analysis
4
Quantification of Cardiac Gene Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Total RNA were extracted from LV tissue with TRIZOL. cDNA was generated from 2μg of total RNA using High Capacity RNA-to-cDNA kit (Life Technology Grand Island, NY) according to the manufacturer’s instruction. qRT-PCR assays were performed using PerfeCT Super Green Super Mix (Quanta Biosciences, Beverly, MA) by using specific primers IL-6, TNF-α, ANP, BNP, β-MHC, Fibronectin, Collagen I, Collagen IV, TGF-β, CTGF, and MMP9. (Integrated DNA Technologies, Inc. Coralville, Iowa). Differences in expression were determined by the relative quantification method; the cycle threshold (CT) values of the target genes were first normalized to the CT values of endogenous control GAPDH.
5
Quantification of Cardiac Gene Expression
6
Quantifying Cardiac Metabolic Gene Expression
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