Read the DNA sequence from one end to the other
& add up the DNA strength parameter (Given in table 1).
↓
Calculate the strength parameter per base (E) as: E = DNA strength parameter ⁄ Length of sequence
↓
Substitute the value of E, the length of sequence
, the Na^{+} ion concentration and the nucleotide strand concentration of the solution in the
given equation Tm (°C) = (7.35 ×E) + [ 17.34 ×ln(Len) ] + [4.96 ×ln(Conc)] + [0.89× ln(DNA)]
 25.42
Figure:Flow chart to calculate the melting temperature
