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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 tool may be cited as follows

MLA

"Quest Calculate™ Temperature of Annealing Calculator." AAT Bioquest, Inc.25 Dec2024https://www.aatbio.com/tools/solve-equation-calculator/temperature-of-annealing.

APA

AAT Bioquest, Inc. (2024December 25). Quest Calculate™ Temperature of Annealing Calculator. AAT Bioquest. https://www.aatbio.com/tools/solve-equation-calculator/temperature-of-annealing.
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