Primer Dimer problems in real-time PCR - (Nov/22/2005 )
Is there any way that the % agarose or voltage or run time is making is hard to discern between primer dimers (which can be 20~80bp, or bigger) and a 100bp PCR rpoduct? Im sure you've addressed this, but just in case...
Also, compare the melt temps of the pos control to the NTC
Different sofwtare give different results and then following the best one in the industry turns out to be the best option. If you have funds available, purchase Beacon Designer from Premier Biosoft. If no try out the free version to check the secondary structures in your primers/probes. It is a free qPCR tool