Annealing Temperature Calculator
Calculate optimal PCR primer annealing temperature based on melting temperature (Tm) calculations.
Primer Sequences
Tm Formulas
Basic: Tm = 2(A+T) + 4(G+C)
Salt: Tm = 81.5 + 16.6×log[Na+] + 0.41(%GC) - 675/N
Optimal Annealing Temperature
46.8°C
Range: 43.8°C - 49.8°C
Forward Primer
Melting Temperature (Tm)54.4°C
Length21 bp
GC Content52.4%
CompositionA:5 T:5 G:6 C:5
Reverse Primer
Melting Temperature (Tm)51.8°C
Length20 bp
GC Content50.0%
CompositionA:5 T:5 G:5 C:5