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: |
Feedback
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This online tool may be cited as follows
MLA | "Quest Calculate™ Temperature of Annealing Calculator." AAT Bioquest, Inc., 25 Dec. 2024, https://www.aatbio.com/tools/solve-equation-calculator/temperature-of-annealing. | |
APA | AAT Bioquest, Inc. (2024, December 25). Quest Calculate™ Temperature of Annealing Calculator. AAT Bioquest. https://www.aatbio.com/tools/solve-equation-calculator/temperature-of-annealing. | |
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