Gene expression was determined by quantitative RT-PCR. Total RNA was extracted and purified from LV tissue and isolated cardiomyocytes with the RNeasy Fibrous Tissue Mini Kit (QIAGEN), following the manufacturer’s protocol. RNA (500 ng) was reverse transcribed into cDNA using the SuperScript III First-Strand Synthesis SuperMix kit (Invitrogen). cDNA was subjected to PCR amplification to detect ANP (Nppa), BNP (Nppb), α-SA (Acta1), collagen III (Col3a1), and Trpm4 gene expression, performed with the CFX384 Touch Real-Time PCR Detection System (Bio-Rad), PCR master mix LightCycler 480 SYBR Green I Master (Invitrogen). Samples were run in technical triplicate, and the mRNA expression levels were normalised to those of GAPDH to calculate relative gene expression using delta-delta Ct method. The mouse RT-PCR primers (Sigma-Aldrich) used are shown in (Key resources table).
Mouse rt pcr primers
Manufactured by Merck Group
Mouse RT-PCR primers are designed for use in reverse transcription-polymerase chain reaction (RT-PCR) experiments involving mouse samples. They are used to amplify and detect specific mouse gene sequences.
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Lab products found in correlation
2 protocols using mouse rt pcr primers
1
Quantitative RT-PCR for Cardiac Gene Expression
2
Gene Expression Profiling in Cardiomyocytes
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Gene expression was determined by quantitative RT-PCR. Total RNA was extracted and purified from LV tissue and isolated cardiomyocytes with the RNeasy Fibrous Tissue Mini Kit (QIAGEN), following the manufacturer's protocol. RNA (500 ng) was reverse transcribed into cDNA using the SuperScript III First-Strand Synthesis SuperMix kit (Invitrogen). cDNA was subjected to PCR amplification to detect ANP (Nppa), BNP (Nppa), α-SA (Acta1), collagen 3 (Col3a1), and Trpm4 gene expression, performed with the CFX384 Touch Real-Time PCR Detection System (Bio-Rad), PCR master mix LightCycler 480 SYBR Green I Master (Invitrogen). Samples were run in technical triplicate and the mRNA expression levels were normalized to those of GAPDH to calculate relative gene expression using delta-delta Ct method. The mouse RT-PCR primers (Sigma-Aldrich) used are shown in Supplementary Tab. 3.
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