TRIzol reagent was used to extract total RNA from cell cultures according to the manufacturer’s instructions. Complementary DNAs were synthesized using PrimeScript RT reagent Kit (TaKaRa, Tokyo, Japan), and real-time PCR were performed by ABI Real-Time System (Applied Biosystems, Foster, CA, USA). The real time PCR primers of genes are listed in Table S1 .
Abi real time system
Manufactured by Thermo Fisher Scientific
Sourced in United States, Canada
The ABI Real-Time System is a laboratory instrument used for real-time quantitative PCR (qPCR) analysis. It is designed to accurately measure and analyze DNA amplification in real-time during the PCR process. The system includes a thermal cycler, optical detection components, and associated software to facilitate quantitative gene expression studies, genotyping, and other real-time PCR applications.
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Lab products found in correlation
2 protocols using abi real time system
1
RNA Extraction and Real-Time PCR Analysis
2
Quantifying MIF gene expression
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After preparing 5 ml of peripheral blood at the beginning and end of the experiment from the patients participating in this study, mRNA extraction was performed using a highly accurate RNA isolation kit (Roche Diagnostics, Germany). Expression of MIF and β-actin genes was measured using the real-time reverse transcriptase (RT-PCR) technique. For sequential amplification of RT-PCR, 2μl of each cDNA sample was used for 20μl of PCR-mix, and PCR reactions were performed in the step of an ABI real-time system (Applied Biosystems, Foster City, Canada).
The β-actin gene sequence, which was used as an internal control, and the MIF gene, was obtained from the NCBI gene bank, and their specific primers were designed by an express primer program. Primer sequences were blasted at NCBI and Gene Runner to confirm the specificity and accuracy of the designed primers. The sequences of primers are listed in Table 1.
The β-actin gene sequence, which was used as an internal control, and the MIF gene, was obtained from the NCBI gene bank, and their specific primers were designed by an express primer program. Primer sequences were blasted at NCBI and Gene Runner to confirm the specificity and accuracy of the designed primers. The sequences of primers are listed in Table 1.
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