PCR Optimization: Finding the Optimal Primer Annealing Temperature

摘要

The base composition and length of PCR primers generally determine the annealing temperature of primer pairs. New primer pairs are designed to anneal at a specific temperature by calculating the primer melting temperature (T_m). The T_m is the temperature, at which one-half of a particular DNA duplex will dissociate and become single strand DNA. The stability of a primer-template DNA duplex can be measured by its T_m and new primers are usually supplied with an additional data sheet providing information on the calculated theoretical melting temperatures (T_m).