"Breathe on Me, Breath of God" has been set to a number of tunes. The most commonly used ones are John Chetham and S.S. Wesley's 1718 "Aylesbury" and blind London organist Charles Lockhart's "Carlisle". Other tunes include "Veni Spiritus" by Sir John Stainer and "Trentham" by Robert Jackson. The use of "Trentham" was criticised by hymnologist Donald Webster in 1980 who stated "One might conclude from [this tune] and the way it is sung that the breath of God was an anaesthetic, not a 'Giver of Life'."
