Real time PCR - no product - (Feb/01/2006 )
I am doing real-time PCR. I designed the primers using Beacon Designer software (Bio-rad), optimised the annealing temperature to 55C. The melt curve is good (no primer dimer,one single peak ). The standard curve,PCR coefficiency was also close to normal.But there is no product on the gel after electrophoresis. I can't figure out where the problem lies.
your suggestions would be of great help.
But what you see in the electrophresis photo?
I had one reaction that the product was right on the same place of the primer´s clowd, and it was difficult to me the product.
How much bp had your product? Use a molecular marker to confirm the bands.