This software can calculate the optimum PCR annealing temperature (Tm value) of the primer.

1. Enter the primer sequence in the textbox for which the Tm value needs to be calculated, and then the primer-related information will be shown below.

2. Click the‘Empty’button to continue calculating the Tm value of another primer.

Note:

The Tm value is calculated by Nearest Neighbor Thermodynamic Theory (NN).

The default calculation parameters are as follows:

Nucleic acid concentration: 250 pM

K+ concentration: 50 mM

Mg++ concentration: 1.5 mM

The Tm value calculated using above parameters are suitable for most PCR reactions without further optimization.