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Combined with the $1 million she gave in 2007, the donation becomes the museum's largest to date. In honor of the media mogul's generosity, a 350-seat theater will be named the Oprah Winfrey Theater.
"I am so proud of African American history and its contributions to our nation as a whole," said Winfrey in a statement. "I am deeply appreciative of those who paved the path for me and all who follow in their footsteps. By investing in this museum, I want to help ensure that we both honor and preserve our culture and history, so that the stories of who we are will live on for generations to come."
Currently under construction on a five-acre site adjacent to the Washington Monument in D.C., the museum is expected to open in late 2015.
Winfrey has been a member of the museum's advisory council since 2004.