Temperature of Annealing Calculator

The annealing temperature is the temperature at which the PCR primers bind to the complementary single-stranded DNA template. It heavily depends on the melting temperature (Tm) of the selected primers. The annealing temperature is usually set a few degrees below the Tm of the primers. Choosing the appropriate annealing temperature is critical for successful PCR amplification as a temperature that is too low can result in non-specific amplification, while a temperature that is too high can result in reduced PCR efficiency.