The Twilight Rune originally resided at the Western Palace, but was removed by orders of Gizel Godwin to battle the Prince's Dawn Rune around the time that Doraat was under Siege. It was equipped to Alenia, with the help of Jeane, however since the rune did not "choose" her, she could not use it to its full potenial. After her defeat and subsequential saving by Dolph, the rune was removed and given to Sialeeds. The rune sensing her royal blood equipped itself to her and she was able to use the rune during the battle of Stormfist. However, in a stunning twist, she used it to kill all of Godwins archers who (along with Childerich) had set a trap for the Prince and his party. The Twilight Rune also played a slight role in the battle against Sialeeds when she revealed to everyone (much to Zerase's dismay) that she had been choosen by the rune. Later after the siege for Sol-Falena, Sialeeds and the Prince had to use the combined powers of the Dawn and Twilight runes to prevent the Sun Rune from destroying all of Raftfleet. Soon afterwards, Lyon was choosen by the rune as Sialeeds died. Later, after victory at Sol-Falena and Godwin stealing the Sun Rune, Lyon and the Prince travel to Lunas to open a secret path to the Ashtwal Mountains under the Lunas Fountain by using their runes. At the Ashtwal Mountians, they discovered more Sindar Ruins, but these are dedicated to the Sun Rune. As with the other ruins, the Dawn Rune and the Twilight Rune work there to open doors. After defeating the Sun Rune Incarnation, Lyon succumbs to her wounds and dies, passing the Twilight Rune onto the Prince. Using the Dawn, Twilight, and Sun Runes, the Prince is able to bring Lyon back to life. Later, after the game is finished, one can see the Dawn, Twilight, and Sun runes in the Sealed Room atop the Sun Palace.
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- Evening Dusk: Causes all enemies to sleep (20%)
- Setting Sun: 300+ damage or instant death (70%) to one enemy
- Silent Afterglow: 600+ damage to row of enemies
- Vermilion Sky: 1300+ damage to all enemies
- During the major/war battles, Twilight Rune can be used to reduce the number of enemy troops, but, because Alenia was never the true master of the rune, it has no effect against the army of the Prince and his allies.
- With the exception of the last power, the Dawn Rune and Twilight Rune generally have opposing powers.
- The first power, Dawn Rune removes status while Twilight Rune causes silence and sleep.
- The second power, Dawn Rune can damage one to three enemies at the same time, while Twilight Rune can only cause damage to one enemy but can cause instant death.
- The third power, Dawn Rune can heal to one ally, while Twilight Rune damage one to three enemies at the same time.
- In the second fight against Sialeeds, she can use Vermilion Sky, the last power of the Twilight Rune, while Freyjadour can not use the maximum power of the Dawn Rune.
- The Twilight Rune holds many allusions to the Black Sword Rune.
- Neither Rune is an actual True Rune, instead they are special runes derived from actual True Runes and thus possess more power than normal runes and a minor will of their own.
- Both Runes focus on offense and hurting the enemy.
- Both runes are paired with an opposing counter rune of similar origin and power.
- Both Runes seem to drain the life force of their user when it's power is overexerted, though in the Twilight Runes case, this is due to being forcibly placed on a non-worthy bearer.