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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.
Equation
Temperature of annealing
=
0.3
×Temperature of primer melting
+
0.7
×Temperature of target melting
-14.9
Inputs
| Temperature of annealing: | |
| Temperature of primer melting: | |
| Temperature of target melting: |
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References
This online resource may be cited as follows
MLA | "Quest Calculate™ Temperature of Annealing Calculator." AAT Bioquest, Inc., 16 Oct. 2025, https://www.aatbio.com/tools/solve-equation-calculator/temperature-of-annealing. | |
APA | AAT Bioquest, Inc. (2025, October 16). Quest Calculate™ Temperature of Annealing Calculator. AAT Bioquest. https://www.aatbio.com/tools/solve-equation-calculator/temperature-of-annealing. | |
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