In recent years, in addition to 2.4 GHz Wi-Fi signals, 5 GHz signals have been introduced making it possible to transmit data at both IEEE 802.11n and 802.11ac standards. Therefore, researchers have increasingly focused on developing indoor positioning applications based on Wi-Fi dual band. This study was conducted in two stages. In the first stage, we investigated the behaviours of 2.4 and 5 GHz Wi-Fi signals by collecting received signal strength (RSS) values from access points and determined the relationship between RSS and distance for each signal frequency using a curve fitting technique. Furthermore, we comparatively analysed signal fluctuations and their effects on positioning accuracy. In the second part of the study, we compared the positioning accuracy of four algorithms; namely, bilateration, trilateration, weighted iterative non-linear least square and extended Kalman filter using 2.4 and 5 GHz Wi-Fi signals. The experimental results revealed that the 5 GHz signals were more stable and had better positioning accuracy than the 2.4 GHz signals. Concerning the positioning algorithms, bilateration had the best positioning accuracy at both frequencies.