This application calculates the Tm for a primer, and gives instructions on how to dilute the primer to a desired concentration.

Important Note:

If the PCR primer contains desired mismatches, e.g. for creating a mutation or a restriction site, make sure to calculate the Tm only for the correctly matched sequence.

Type or paste your sequence

5`-

PCR Conditions

Primer conc. µM

Salt conc. mM

Result

Tm °C (Modified Breslauer's thermodynamics, dH and dS parameters)

Tm's according to other methods

Tm ^oC (Allawi's thermodynamics, dH and dS parameters)

Tm ^oC

Molecular weight g/mol

Extinction coeficient l/(mol˙cm)

Primer dilution

Amount of primer

OD₂₆₀

Mass µg

nmol

Preparing stock solution

Desired stock concentration µM

Dissolve the primer in µl of H₂O or buffer

Dilution

Desired volume µl

Desired concentration µM

Dissolve µl of the stock solution

Volume of the solvent µl of H₂O or buffer