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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