Dear molecular biologist experts,

I am combining three probes that should be revealed in a multiplex real time pcr with the system LC480. Do you have any experience with dramatic drop down of fluorescence in a triplex with the following probes and channel?

Channel:Exc-Det

FAM : 465-510

LC610 : 533-610

CY5 : 618-660

protocol:

(95°C 5 min) x 1 cycle

(95°C x 10 sec - 60°C x 45 sec - 72°C x 1 sec) x 40 cycles

(40°C 10 sec)

Let me know what do you think about