Customers often ask "How do I determine the best annealing temperature (Ta) for my primers?"
To determine the best annealing temperature for the primers using the site-directed mutagenesis kit, we recommend that you use the NEBbase changer tool. The NEBbase changer tool takes into account the mismatches in your mutagenic primer set. Standard Tm calculators do not take into account those mismatches, so the Tm that you get from those tools may not be optimum.
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