It is recommended that a calculation of the optimum annealing temperature (Ta) for a primer be performed using one of several empirical formulas. A rough calculation can be made by adding 2°C for each A and T nucleotide present in the primer and 4°C for each G and C nucleotide and subtracting 5°C from the result.
Ta = Tm - 5°C
Maar hoeveel is Tm??