There are two algorithms for melting temperature calculation:
- Rough
- Primer 3
Rough
The melting temperature is calculated as follows. For sequences of length 15 or longer:
Tm = 64.9 + 41 * (nG + nC - 16.4) / (nA + nT + nG + nC) |
For shorter sequences:
Tm = (nA + nT) * 2 + (nG + nC) * 4 |
Here "nA", "nT", "nC", "nG" denote the number of the corresponding nucleotide.