Conventinal PCR works but qPCR does not

[flyman]

Hello, I tried several times to run a RT-PCR that has not yet worked. I know that the primers, polymerase, and especially the template are fine as I have done regular PCR many times and seen the exact amplifide product that I should be seeing. However, whenever I use the RT-PCR machine and it fails, I run the rxn out on a gel to find nothing but probable primer dimers??Again there is no evidence of Primer dimers in conventinal PCR with the same amount of template. Do you think it is the machine. What factors could cause an RT-PCR rxn to not work but a regular PCR does work?

BTW: PCR conditions are exactly the same between the two machines with the exception of the elongation step. It is 15 seconds longer on the pPCR to allow for better detection...

Thank you for your insightful comments

[danfive]

First of all What is your detection probe? SYBR Green Dye, Taqman?
Are the primers the same ones for qPCR as for normal PCR?
Do you have a known working qPCR assay, primers and controls for B-actin (or whatever) to test machine?
Lastly, How many PCR cycles in normal PCR and qPCR? What concentration are the DNA templates and primers/probe?