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Quantitative Determination of IFN-γ mRNA by the Competitive RT-PCR Method

Zhu Qinghui,Shuai Yuemin,Zhang Gusheng (Shanghai Chang Jiang Center for Immunological and Genetic Laboratory Technology,First Hospital of Shanghai Textile,Shanghai 200060)  
To explore the optimal conditions for quantitative determination of IFN γ mRNA by the competitive RT PCR method,the internal standard of cDNA fragment was prepared and used as the competitive templet The IFN γ mRNA in PBMC of healthy volunteers induced by 2 0μmol/L of calcium ionophore and varying concentrations of PMA was determined by the established method The experimental results showed that the expression of IFN γ mRNA in the healthy volunteers stimulated with ionophore and PMA harvested at 6 hours reached the highest level This method is simple and sensitive,and can be used for the quantitative determination of cellular IFN γ mRNA and for the studies of mechanism of IFN γ genes expression
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