Enter the melting temperature of the primer and the melting temperature of the product to determine the optimal annealing temperature.

## Annealing Temperature Formula

The following formula is used to calculate the optimal annealing temperature of a PCR

AT = (.3 * Tm primer ) + ( .7 * Tm product) – 14.9

- Where AT is the optimal annealing temperature
- Tm primer is the melting point of the primer
- Tm product is the melting point of the product.

## Example

Let’s take a look at an example of how to calculate the annealing temperature.

- First, we must determine the melting temperature of the primer. For this example we will assume the temperature is 50 C.
- Next, we must determine the melting temperature of the product. For this example we will assume that is 75C.
- Finally, we can calculate the annealing temperature using the formula above and we yield as results of an optimal temperature of 52.60 C.

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