Nucleic acid hybridization melting temperature

Tm is the melting temperature of a double-stranded nucleic acid. If the melting temperatures of the primers are too different, the extent of hybridization with the target DNA will differ during the annealing phase, which would result in differential replications of the strands. 

The melting temperature (T ) of nucleic acids defines the thermal point at which 50% of the complementary molecule dissociate/melt away from their cognate strands or anneal/hybridize the two strands. The capacity to estimate melting temperatures is important because it helps the researcher select strand separation/hybridization temperatures, such as in polymerase chain reaction and microarray assays. A good approximation of melting tonperature for oligonucleotides takes the expression (Wallance et al., 1979) ... 

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