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Tallis began his career as organist at the Benedictine Priory at Dover, followed by similar service at Waltham Abbey until the dissolution of the monasteries in 1540. He was then organist at Canterbury Cathedral and in 1543 became a Gentleman of the Chapel Royal, a position he retained until his death. Like Byrd, his pupil, he seems to have remained loyal to the old religion, while continuing to enjoy royal favor.

Tallis wrote a quantity of Latin church music and contributed also to the reformed English liturgy, in some cases adapting earlier Latin compositions. One of his most remarkable achievements is the 40-voice Spem in alium. His setting of the Latin Holy Week liturgy Lamentations represents his work at it's height. The composer's name is widely known through the metrical psalm tune known as Tallis's Canon, a setting of God, grant we grace.