Primer Dimer problems in real-time PCR

[moleculardude]

I am having primer dimer problems in real-time PCR...

what tips do you guys have in fixing this. I even ordered the primers from a company they were supposed to be good!

[Leowe]

I even use primer design software, what results Self Dimer dG
Mb u need to order/choose another primers?

Another way is:
- Set anneling time to minimun
- set concentration of primers to minimum
- increase Ta

or if u detect dimers only in low concentrations of substract u can try to add incubation b4 Plate Read just around 1-2 sec (Temperature: hi then T denaturation of dimers, but lower then T denaturation of ur product)

This information was taken form molbiol.ru
Sorry for my bad english

[waleedkars]

I first suggest that the primer concentration be set to a minimum after adjusting for optimal primer:target sequence ratio. I also suggest that one need to re-design the primers such that the last three NTDs at 3' end contain no G or Cs. Increasing the annealing temperature will not help much if the primer:target ratio is far from optimum

[kiki06]

Yes, those are very good points.

I would ask for the sequences as mentioned and check them in a primer design program for self-annealing!

[lenie_86]

make a hot start and do not allow to stand your real time mix + template at room
temperature.

[attc79]

Thanks