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We don't know when River (Melody Pond) first learned that Amy was her mother. It appears, however, that she already knew it by the time she was in the orphanage in 1969 Florida ("Day of the Moon"). In her room in the orphanage, she had photographs of Amy holding her as a baby. Those photos were presumably taken on Demons Run ("A Good Man Goes to War"), since that's the only place we know of where the two of them were together while River (Melody) was a baby.