Total RNA was extracted from 3D4/21 cells using the TRIzol reagent (Invitrogen) according to the manufacturer’s instructions, and the cDNA synthesis was performed using the PrimeScript reverse transcriptase reagent kit (TaKaRa, Dalian, China). Quantitative PCR analysis was performed using TB Green Premix Ex Taq (TaKaRa) on an ABI QuantStudio 5 real-time PCR system (Agilent Technologies, Santa Clara, CA). The relative mRNA level of each gene was normalized to GAPDH mRNA levels and determined based on the standard 2–ΔΔCT protocol. The primers used for RT-qPCR were listed in Table 1 .
Abi quantstudio 5 real time pcr system
Manufactured by Agilent Technologies
The ABI QuantStudio 5 real-time PCR system is a laboratory instrument designed for real-time polymerase chain reaction (PCR) analysis. It is capable of performing quantitative PCR (qPCR) experiments to detect and quantify specific DNA or RNA sequences.
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2 protocols using abi quantstudio 5 real time pcr system
1
RNA Extraction and qPCR Analysis
2
RNA Extraction and qPCR Analysis
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Total RNA was extracted from 3D4/21 cells using the TRIzol reagent (Invitrogen) according to the manufacturer’s instructions, and the cDNA synthesis was performed using the PrimeScript reverse transcriptase reagent kit (TaKaRa, Dalian, China). Quantitative PCR analysis was performed using TB Green Premix Ex Taq (TaKaRa) on an ABI QuantStudio 5 real-time PCR system (Agilent Technologies, Santa Clara, CA). The relative mRNA level of each gene was normalized to GAPDH mRNA levels and determined based on the standard 2–ΔΔCT protocol. The primers used for RT-qPCR were listed in Table 1 .
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