The RNA was extracted from tissues by using Trizol Isolation Reagent (#79306; Qiagen, NRW, Düsseldorf, Germany) according to the manufacturer’s protocol. Additionally, then, the RNA was reverse transcripted into cDNA using Revers Transcription-premix (#EBT-1514; Elpis Biotech, Daejeon, Republic of Korea). The cDNAs were used for qRT-PCR analysis with primer sets in Table 1 and Table 2 , and SYBR Green (#RT501; Enzynomics, Daejeon, Republic of Korea), according to the manufacturer’s protocol. Finally, the gene expression of cDNA each group used the StepOnePlus Real-Time PCR systems (Applied Biosystems, Austin, TX, USA). Gene expression was normalized to the Ct values of GAPDH in each sample and was calculated by the 2−ΔΔCt methods.
Trizol isolation reagent
Manufactured by Qiagen
Sourced in Germany
Trizol Isolation Reagent is a single-step RNA isolation solution that facilitates the extraction and purification of total RNA from various biological samples, including cells, tissues, and microorganisms. It is a phenol-based reagent that effectively lyses the samples and separates the RNA from DNA and proteins during the isolation process.
Automatically generated - may contain errors
2 protocols using trizol isolation reagent
1
Quantitative Analysis of Gene Expression
2
Quantitative Analysis of mRNA Levels
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Total RNA from MLE-12 cells was isolated using Trizol Isolation Reagent (#79306; Qiagen, NRW, Düsseldorf, Germany). For reverse transcription-polymerase chain reaction, 1 µg of total RNA was reverse transcribed for cDNA synthesis in each tube using Revers Transcription-premix (#EBT-1514; Elpis Biotech, Daejeon, Korea). Quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) was used with SYBR Green (#RT501; Enzynomics, Daejeon, Korea) for gene expression of mRNA levels. Two pairs of primers are shown in Table 1 . Relative quantification of gene expression was normalized Ct values of β-actin each sample and calculated by 2−ΔΔCt methods.
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