so i am stuck...i would like to do quantitative PCR rather than qualitative, and have been unable to get the quant. to work.
in theory, there should be no difference, except for the addition of the fluorescent dye to the mix for teh real-time.
for the regular PCR i use amplitaq gold. for real-time, i use AB's 2X sybr master mix. the regular PCR works fine, it's never a problem. but i tried moving on to the real-time and i get crazy results, with lines all over the place when i try to look at the amplification curves (it's an abi7300 machine). i tried using new samples in case that was the problem, i tried using just total genomic DNA, same craziness. i ran the samples out on a gel, and not suprisingly, no band.