It is compatible with the On Tour Guitar Grip, as well as a unique "Drum Skin" and microphone support, but is not considered part of the On Tour series.

The game recognizes strums in any direction as well as back-forth strums without lifting up the pick, a technique that comes in extremely handy when you get into the Expert tracked songs. The theme song was released as a free download for Guitar Hero World Tour the week before the game's release.Game Informer stated that, compared to the setlists for other Guitar Hero games, the songs "skew away from great guitar rock and into the territory of pop/rock", and "just aren't as exciting to play". Players use the Guitar Hero pick-stylus on the touch screen to strum their favorite tunes and create their rock 'n' roll legacy in career mode. Guitar Hero: On Tour features two different setlists, one for primarily English-speaking regions including North America, the British Isles, and Australia, in addition to Japan and the Netherlands, [4] and another for other non-English-speaking European countries, with five replacement songs; [5] songs not in the game for that region are marked as "N/A" in the table below.

The local wi-fi capabilities of the DS are used for multiplayer mode, and allow a player to use songs from one installment of the series in competitive modes with a player with a different installment.There is a single player Career Mode, allowing the player to select from new characters introduced for On Tour or previous characters from the other games in the Guitar Hero series. Other requests are based on the effects players use in multiplayer mode, such as playing an entire song at "Hyperspeed" (notes moving on the screen faster than normal) or by using the whammy bar on every sustained note. Each song has been divided into three separate compressed tracks that play back simultaneously during gameplay. To start the purchasing process, it is necessary to sign in with the Nintendo Account and the Nintendo Network ID.