This is an empirical input-Tm estimate, not a sequence-design or thermocycler protocol generator.
How to use Annealing Temperature Calculator
- Read the method and enter quantities in the units shown on the form.
- Check the measurement basis, recorded event, genetic assumptions or the externally supplied care-plan instruction before calculating.
- Select Calculate, then read both the result and its limitations. Reset clears the result and restores the starting form.
Method and scope
Ta ≈ 0.3 × lower primer Tm + 0.7 × product Tm − 14.9 °C.
Work through the input issue
For a related perspective, read Primer Tm and Annealing Temperature: Keep the Assay Conditions Together.
Related measurement
qPCR Efficiency Calculator can help with a related question, using its own stated assumptions.
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