7500 fast real time pcr platform

Quantitative real-time PCR (qRT-PCR) was conducted to quantify gene expressions. The RNA samples used for qRT-PCR were the same as with those used for the transcriptome sequencing. The experiments were performed on Applied Biosystems 7500 Fast Real-Time PCR platform with the SYBR^® Premix Ex Taq™ II mix (Takara Biotechnology Co., Ltd, Dalian, Japan), and the results were analyzed by the Applied Biosystems 7500 software (Applied Biosystems Life Technologies). The results of triplicate assays were then calculated by the 2^-ΔΔCt method. The primers for qRT-PCR analysis are listed in Table S2.

The samples collected at 6:00, 12:00, 18:00, and 0:00 h during the full blossoming period were subjected to qRT-PCR tests to determine the transcript abundance of the genes involved in formation of terpenoids. The experiments were performed on Applied Biosystems 7500 Fast Real-Time PCR platform using the SYBR^® Premix Ex Taq^TM II mix (Takara Biotechnology Co., Ltd., Dalian, Japan) and the results were analyzed by the Applied Biosystems 7500 software (Applied Biosystems Life Technologies). Three biological replicates were tested and relative transcript levels were calculated by the 2^-ΔΔC_T method using β-Actin as the endogenous control gene for data normalization. The relative gene expression was determined as previously described by Livak and Schmittgen (2001) (link). The primers for qRT-PCR analysis are listed in Supplementary Table S1.