How were the primer melting temperatures calculated?
The primer Tm values are calculated using the Nearest Neighbor Method with the up-to-date thermodynamic parameters. Tm values are also dependent on primer and salt concentrations. Higher concentrations of primer and salt usually lead to higher Tm. The Tm values included in PrimerBank are for 0.25 uM primer, 1.5 mM Mg2+, 50 mM Na+, and 0.8 mM dNTP, which are typically used in PCR experiments. Other common PCR conditions only affect the Tm slightly.
