What is the minimal concentration of PCR primers for a successful PCR reaction? Let's do this simple experiemnt to have template concentration fixed, TaqMan probe concentration fixed, all PCR setting constant, only change primers concentration to see the effects of Ct outcome.

Fig. 1000 copies of DNA templates were PCR amplified in 20uL, with 300nM constant probe concentration and varying forward and reverse primer concentrations. PCR annealing time was fixed at 1 minute. Results revealed the practical minimal PCR primer concentration is about 300nM.