Figure 1. (A) The fluorescence signal changes of MYCN MB in the presence and absence of the target sequence by increasing the temperature from 25 °C to 95 °C. Inset: the fluorescence signal fold change of MYCNMB varies from 25 °C to 95 °C in the presence and absence target sequence. (B) The fluorescence signal changes of NAGK MB in the presence and absence of the target sequence by increasing the temperature from 25 °C to 95 °C. Inset: the fluorescence signal fold change of NAGK MB varies from 25 °C to 95 °C in the presence and absence of target sequence. (C) The relationship between the fluorescence value of MYCN MB and the target sequence. Inset: calibration curve of MYCN MB fluorescence value with the target sequence. (D) The relationship between the fluorescence value ofNAGK MB and the target sequence. Inset: calibration curve ofNAGK MB fluorescence value with the target sequence. (E) Real-time fluorescence curves of the response of the developed single tube duplex RT-PCR to different copy numbers of MYCN . Inset: theC _T values of the developed single tube duplex RT-PCR show a logarithmic linear correlation with the MYCN copy number. (F) Real-time fluorescence curves of the developed single tube duplex RT-PCR for NAGK response with different copy numbers. Inset: The C _T values of the developed single tube duplex RT-PCR show a logarithmic linear correlation with the NAGKcopy number.