The funds will support efforts by the United Nations High Commissioner for Refugees to provide "lifesaving protection and assistance to those fleeing the conflict," National Security Council spokesman Tommy Vietor said in a statement.
Vietor said nearly 230,000 Malian refugees have fled to Algeria, Burkina Faso, Mauritania and Niger, while an additional 155,000 Malians have been displaced within their country.
"The United States is deeply concerned about the situation of the Malian people," Vietor said. "We call on all parties to support the restoration of democratically elected civilian governance in Mali as soon as possible. Specifically, we ask that the interim government issue its road map for elections without delay so that preparations can begin in earnest."
He called on the military-led National Committee for the Restoration of Democracy not to interfere in political matters.
Vietor said the United States strongly condemns attacks against civilians in northern Mali and the reported destruction and looting of religious, historical and cultural sites in Timbuktu.
"We call on the rebel groups in northern Mali to renounce any connection with terrorist groups and enter into legitimate political negotiations," he said.
The United States also is calling on "all parties to ensure neutral, impartial and unhindered humanitarian access to all populations in northern Mali," Vietor said.
He also said the United States is continuing to support the leadership of the Economic Community of West African States in its mediation efforts "and will continue to consult with ECOWAS and other regional stakeholders on the best way to facilitate the political transition and restore peace and security across Mali."