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

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.