2. Properties of DNA

2.4 Tm calculation for a DNA molecule

Tm is the temperature at which one half of a DNA molecule is denatured.

The bond between the AT bases is weaker (because there are two hydrogen bonds) than the bond between GC bases (because there are three hydrogen bonds).

A Tm calculation must take into account:

  • the number of matching AT bases (= 2°C for each base pair)

  • the number of matching GC bases (= 4°C for each base pair)

Tm= 2(AT) + 4(GC)

  • AT = 2°C

  • GC = 4°C

This formula is restricted to small-sized DNA molecules (between 17 and 25 base pairs).