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An effective methodology is proposed to evaluate the boundary stiffnesses of beams with generally restrained edges. The boundary conditions at the ends of the beams are implemented by the penalty method. The unified Jacobi polynomials are introduced to represent the assumed mode shape functions of the beam. Hamilton’s principle with the assumed mode method is employed to derive the equation of motion of the beam with generally restrained boundaries. Subsequently, a novel approach is proposed to evaluate the boundary stiffnesses of the beam using the measured natural frequencies of the beam by vibration experiments. The accuracy of this method is verified by comparing the results with the previously published literature, the finite element method (FEM), and the vibration experiments. The effects of the types and values of the boundary spring stiffnesses on the vibration properties of the beam are also discussed. The calculation method in this research can provide an effective prediction technique for the boundary stiffnesses of engineering structures with generally restrained edges. [ABSTRACT FROM AUTHOR]