I'm testing fluorescence level of a sample having dsDNA, SYBR Green and milli q water but I'm facing difficulty in choosing right volume of dsDNA and cons level of SYBR Green.

Details of sample: dsDNA:1000 nano mole, SYBR Green:1X (1000 times diluted from original stock)

Sample_1: dsDNA (3ul) + SYBR Green (5ul) + milli q water (27ul)

Sample_2: dsDNA (3ul) + SYBR Green (5ul)

Result: In first sample I haven't got any signal but in second case I got( But I want 5 times higher then what I got).

Please tell me how it work and how these things depends on each other. I'm not a biologist if possible suggest me resource as well.