What is the optimal annealing temperature for my oligonucleotide pair?
The optimal annealing temperature (OAT) is the temperature where PCR of a given fragment is successful. This temperature varies depending on the primers and the melting temperature of the entire fragment. Empirically OAT is normally between 5 and 10 degrees Centigrade lower than the primer to target Tm of the lowest Tm primer in a pair. This empirical observation gives a good starting annealing temperature (T A ) for amplification reactions. Contact Us if you have questions not answered here.
