Miriam Margolyes was born in Oxford, England, on 18 May 1941, the only child of Ruth (née Walters; 1905–1974), a property investor and developer, and Joseph Margolyes (1899–1995), a physician from Glasgow. She grew up in a Jewish family; her ancestors migrated to the UK from Poland and Belarus. Her great-grandfather, Symeon Sandmann, was born in the town of Margonin in central-western Poland, which Margolyes visited in 2013. She attended Newnham College, Cambridge, where she read English. There, in her twenties, she began acting and appeared in productions by the Cambridge Footlights comedy troupe. Margolyes is a lesbian. On becoming an Australian citizen, on Australia Day 2013, Margolyes referred to herself as a "dyke" live on national television and in front of the then prime minister, Julia Gillard. Since 1967, her partner has been Heather Sutherland a retired Australian Professor of Indonesian Studies. Formerly based in Amsterdam, Margolyes divides her time between homes in London, Tuscany and Robertson, New South Wales. Margolyes is a humanitarian advocate. She is a Palestinian human-rights activist, having been a member of the British-based ENOUGH! coalition that seeks "a just settlement between Israelis and Palestinians". She is also a signatory of Jews for Justice for Palestinians. Margolyes is a campaigner for a respite care charity, Crossroads. In August 2015, she was a signatory to a letter criticising The Jewish Chronicle's reporting of Jeremy Corbyn's association with alleged antisemites. Author and comedian David Walliams says he used Margolyes as a model for the title character in his children's book Awful Auntie after a rude exchange with the actress during a stage production. He stresses that he has nothing against Margolyes and is a fan of her work.

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