水稻GA20ox-2基因mRNA的TaqMan荧光定量RT-PCR检测

A technique for real-time quantification of GA20ox-2 expression in rice using TaqMan fluorescence quantitative PCR with specific primers and probe was established. The preparation of standard plasmid DNA for real-time quantification of transcripts of GA20ox-2 had a good practicability in the established system. The technique was exact, authentic and convenient. Standard curve showed the established system had a strict specificity and sensitivity, and had a 10^2 to 10^7 copies respondent capability of initiative templates, and had a good stability and repeatability, with the coefficients of variation in intra-and inter-batch were 0.12% to 0.31% and 0.21% to 0.34%, respectively. There were a high PCR efficiency(E=100.3%) and a good linear relationship between threshold cycle value at which sample crosses threshold and the logarithmic value of template concentration(correlation coefficient = 0.999). A series of standards for real-time PCR analysis have been constructed successfully, and real-time TaqMan-fluorescence quantitative RT-PCR is reliable to quantitatively evaluate mRNA of GA20ox-2 in rice.