Elongation Temperature and Time
Denaturing Temperature and Time. ... (G+C) content: a simple formula for calculation of the Tm is. Tm = 4(G + C) + 2(A + T)°C. Thus, the annealing temperature chosen for a PCR depends directly on length and composition of the primer(s). One should aim at using an annealing temperature (Ta) about 5°C below the lowest Tm of the primers to be used ................
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