Transmission spectra of the lowest-order antisymmetric (A0) Lamb wave for adhesively bonded single lap joints are experimentally investigated in a frequency range below 0.4 MHz. Based on the theoretical results of the wave interference, a tangential bond stiffness evaluation method is proposed for the lap joints. Aluminum alloy plates bonded with epoxy adhesive on different conditions were prepared as bonded specimens. An air-coupled ultrasonic transducer was used to generate the A0 mode in the specimens, and the signals of the transmitted waves across the joints were measured with a pin-type piezoelectric transducer. Spectral analysis was performed for the measured waveforms, and the transmission coefficient was calculated for each specimen as a function of frequency. The experimental results showed that the transmission coefficient has local maxima and minima at multiple frequencies, called peak and notch frequencies, respectively. The sets of the peak and notch frequencies were different among the bonded specimens. Based on the peak and notch frequencies, the tangential bond stiffness was estimated for each bonded specimen. As a result, the bond stiffnesses obtained for the specimens with the same nominal bond thickness and adherend pretreatment were almost equal even if the bond lengths were different. Furthermore, the evaluation results showed that pretreatments such as sanding and contamination of adherend surfaces affect the bond stiffness due to the change in the interfacial stiffness.