实时荧光定量PCR反应体系的优化研究

Factor optimization for the reaction of real-time fluorescent quantitative PCR

摘要: 为了降低实时荧光定量PCR（Real-Time Fluorescent Quantitative PCR，FQ-PCR）技术的实验成本袁，高利用效率，对影响FQ-PCR较敏感的因素二甲基亚砜（DMSO）、甘油和Mg2+等进行了优化试验。结果表明，10 μL反应体系的最优组分为：1*rTaqBuffer袁0.16 mmol/L dNTPs，0.24 μmol/L Primer，2.5 mmol/L Mg2+，5%甘油，5% DMSO，1*SYBR Green I，0.8 UrTaq 酶，100 ng模板。该反应体系为经济型FQ-PCR的广泛应用奠定了基础。

Abstract: Real-time fluorescent quantitative PCR (FQ-PCR) has been widely applied in biological research. To reduce the cost of experiment and increase the used efficiency, the concentrations of the sensitivity factors including DMSO, Glycerin and Mg2 + were further optimized in the FQ-PCR protocol. The results showed that the best optimal system (10 μL) was as follows: 1*rTaq Buffer, 0.16 mmol/L dNTPs, 0.24 μmol/L Primer, 2.5 mmol/L Mg2+, 5% Glycerin, 5% DMSO, 1伊SYBR Green I, 0.8 U rTaq and 100 ng template. The optimized system was the basis of economical application of FQ-PCR.