The annealing temperature (Ta) chosen for PCR relies directly on length and composition of the primers. Generally, you should use an annealing temperature about 5°C below the Tm of your primers. Conversely, too high a Ta may reduce reaction efficiency, as the likelihood of primer annealing is reduced significantly. Make use of this online annealing temperature calculator to calculate the annealing temperature from the values of Oligo sequence.
The optimal annealing temperature (Ta) is the range of temperatures where the efficiency of PCR amplification is maximal.This annealing temperature calculator helps you to calculate such optimal Ta.