Primer Sequences
Forward Primer (5' to 3')
Reverse Primer (5' to 3') (optional)
Reaction Conditions
Monovalent Cation Concentration [Na+] (mM)
Primer Concentration (nM)
Mg2+ Concentration (mM)
Total dNTP Concentration (mM)
Calculation Method
Nearest-Neighbor (recommended for primers 15-70 nt)
Salt-Adjusted (general purpose)
Basic / Wallace Rule (primers under 14 nt)
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Temperature Unit
Celsius (°C)
Fahrenheit (°F)
Annealing Offset below Tm (°C)
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