Annealing Temperature Calculator

In general, the annealing temperature is a temperature at which the primers bind to the complementary region on the target DNA. It depends on the length of primers, GC content and specificity.

The annealing temperature is a temperature which is lower than the melting temperature which facilities primer binding.

The formula to find annealing temperature is here.

T_a = 0.3 x T_m^p + 0.7 x T_m^t - 14.9

You can use Annealing Temperature Formula and substitute values in it to get the answer.

•  Enter primer melting temperature, target melting temperature details as inputs.
• Hit the calculate button.
• The tool will display the annealing temperature as answer.

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1. What does annealing temperature depend on?

The annealing temperature depends on primer melting temperature and target melting temperature.

2. What is the difference between annealing temperature and melting temperature?

The annealing temperature is the temperature where primers successfully bind. Melting temperature is the temperature where primers get fall off from DNA.

3. What is PCR?

PCR acronym is a polymerase chain reaction which is the most groundbreaking technique in biology. The main principle of PCR is to reduce the replication process of DNA in a vial and it allows to repeat several times.

4. How to find annealing temperature in PCR?

The annealing temperature of the primers in a PCR cycle can be calculated with less stable primer, melting temperature of target DNA. The annealing temperature formula is T_a = 0.3 x T_m^p + 0.7 x T_m^t - 14.9.