Dear Vahhab,

I assume you are diluting your mix to 1x and use no other fluorophores in your mix. Which channel (and in which step) are you reading? I guess it is a Rotogene 6, if you could paste an image about the cycling parameters, that might be helpful. Also, DNA amount looks quite high, did you quantify it before the run?

I'm trying to do with positive control and NTCC control, and then paste new image, i think maybe my DNA quality was low