RNA from cardiomyocytes was isolated using an RNeasy Mini kit (Qiagen), as described in the kit manual. RNA from the cardiac tissue was isolated using TRI reagent (Sigma, St. Louis, MO, USA). RNA samples were treated with RNase-free DNase (Qiagen) to remove DNA contamination. cDNAs were synthesized with 500 ng RNA using a SuperScript III First-Strand Synthesis SuperMix reagent kit (Thermo Fisher Scientific, Waltham, Hercules, California MA, USA). Gene expression was analyzed by qRT-PCR (QuantStudio 7 Flex), using SYBR Green master mix (Thermo Fisher Scientific) with gene-specific primers. Data were normalized with GAPDH or Actin as an internal control.
Superscript 3 first strand synthesis supermix reagent kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The SuperScript III First-Strand Synthesis SuperMix reagent kit is a lab equipment product designed for cDNA synthesis. It includes a reverse transcriptase enzyme, reaction buffer, and other components required for the first-strand cDNA synthesis process.
Automatically generated - may contain errors
2 protocols using superscript 3 first strand synthesis supermix reagent kit
1
Cardiac mRNA Isolation and Quantification
2
Cardiac Gene Expression Analysis
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RNA from cardiomyocytes was isolated using the RNeasy Mini kit (Qiagen). TRI reagent (Sigma, St. Louis, MO) was used for RNA isolation from clinical tissue samples and treated with RNase free DNAse (Qiagen) to remove DNA contamination. cDNA was synthesized with 500 ng RNA using a SuperScript III First-Strand Synthesis SuperMix reagent kit (ThermoFisher Scientific). Gene expression was analyzed by qRT-PCR using SYBR Green master mix (Applied Biosystems) using gene specific primers (Table 1 ). Data were normalized with GAPDH as internal control.
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