How to calculate primer annealing temperature? 💡 Please give the step and suitable formula (manual calculation)? ❓ In my experiment DNA template will used and primer basepair between 18 to 25 basepair (Primer minimum size = 18 and Primer maximum size is 25). thanks for answer. 🙄
I think, pDRAW32 program can calculate it 😉
On synthesis report I has found formula for primers of 13-50bp long:
Tm = 100.5 + (41*(yG+zC)/(wA+xT+yG+zC)) – (820/(wA+xT+yG+zC)) + 16.6*log10[Na+]
The last part adjusts the Tm for changes in the salt conc.