At the indicated time points per experiment, total RNA was extracted using TRIzol reagent (Thermo Fisher) and reverse-transcribed using the PrimeScript RT Reagent kit (Takara) according to the manufacturer’s instructions. The CFX384 instrument (Bio-Rad) with SYBR green fast mixture (Bio-Rad) was used for quantitative real-time PCR analysis. Expression levels were quantified using the 2−ΔΔCt method and normalized to GAPDH. The primer sequences are listed in Supplementary Table 1
Sybr green fast mixture
Manufactured by Bio-Rad
Sourced in United States
SYBR green fast mixture is a ready-to-use solution for real-time PCR applications. It contains SYBR Green I dye, which binds to double-stranded DNA and emits fluorescence upon excitation, allowing for the detection and quantification of DNA targets during the PCR process.
Automatically generated - may contain errors
2 protocols using sybr green fast mixture
1
Quantitative PCR and Cytokine Analysis
2
RNA Extraction and qPCR Analysis
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Total RNA was extracted from DF-1 cells at the indicated time points using TRIzol reagent (TaKaRa, Kusatsu, Japan) and reversely transcribed to cDNA using a PrimeScript RT Reagent kit (TaKaRa, Kusatsu, Japan). qPCR analysis was performed on a CFX96 PCR system (Bio-Rad) using SYBR green fast mixture (Bio-Rad, Hercules, CA, USA). Expression levels were quantified using the 2 −ΔΔC(T) method and normalized to GAPDH. Primers used for RT-qPCR are listed in Supplementary Table 1
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