PCR Annealing Temperature Calculator

Determine optimal annealing temperature from primer Tm values

Primer Tm Values

Forward Primer Tm (°C)

Reverse Primer Tm (°C)

Calculation Method

Annealing Temperature (Ta)

54.0°C

Average Tm - 5°C

Tm Analysis

Average Tm59.0°C

Lower Tm58.0°C

Higher Tm60.0°C

Tm Difference2.0°C

All Ta Methods

Average - 5°C54.0°C

Lower - 5°C53.0°C

Optimal55.0°C

Touchdown Start63.0°C

Gradient PCR Range

48.0°C → 65.0°C