How to calculate primer annealing temperature?

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    • #11891
      veniesh
      Participant

      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. 🙄

    • #93161
      JackBean
      Participant

      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.

      Try http://www.metabion.com/biocalc 😉

      A don’t double your posts!

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