Born Dorothy Crowfoot in Cairo, Egypt, she was educated at Somerville College, Oxford. After a brief period as a postgraduate student at Cambridge University, she returned to Oxford in 1934 and spent her entire academic career there. She served as Wolfson Research Professor of the Royal Society from 1960 until 1977.

Hodgkin had the good fortune to fall under the influence of the inspiring and scientifically imaginative physicist J. D. Bernal at Cambridge. Bernal was keen to use the technique of x-ray diffraction analysis, introduced by Max von Laue in 1912, to investigate important complex organic molecules. He gathered around him a group of enthusiastic scientists to work out the appropriate techniques. Of the Bernal group, Hodgkin was probably the most talented; she also possessed a greater single mindedness than Bernal himself and, despite the demands of three young children and a busy political life, it was her persistence and talent that produced some of the first great successes of x-ray analysis.

Her first major result came in 1949 when, with Charles Bunn, she published the three-dimensional structure of penicillin. This was followed by the structure of vitamin B_{1 2} (by 1956) and, in 1969, that of insulin. For her work on vitamin B_{1 2} she was awarded the Nobel Prize for chemistry in 1964.