He matriculated as a pensioner of St John's College, Cambridge, in December 1576, and on 5 November 1579 was admitted a scholar.
He also preached before Henry Frederick, Prince of Wales at Greenwich on 12 March 1605, and before the kings of England and Denmark at Theobalds, then the residence of Robert Cecil, 1st Earl of Salisbury, on 27 July 1606.
On 4 November 1602 Chamberlain had written to Carleton that "Dr. Plafer, the divinity reader, is crazed for love", and after 1606 Playfere's mind gave way, but he held his professorship until his death, on 2 February 1609.
His funeral sermon was preached by Thomas Jegon, vice-chancellor; John Williams made an oration on him in the college chapel.
He was buried in the St Botolph's Church, Cambridge, where a monument with his bust, and a panegyrical inscription, was placed by desire of his wife Alicia.