Jerome Karle was born in Brooklyn, New York, on June 18, 1918, to Louis Karfunkle and Sadie Kun. He attended Abraham Lincoln High School in Brooklyn from 1929 to 1933. Upon graduating, he enrolled in chemistry and biology at the City College of New York. In 1937, he was awarded his B.S. degree and the First Caduceus Award for Excellence in the Natural Sciences. He then studied biology for a year at Harvard and received his M.A. in 1938. Karle worked as a laboratory assistant at the New York State Health Department in Albany from 1939 to 1940. One of his discoveries, a method of measuring the amount of fluorine in water, became a standard in the field.

In 1940, Karle enrolled as a Ph.D student in the chemistry department of the University of Michigan. A classmate of his, Isabella Lugoski, was a fellow chemist whom he would marry in 1942; they would have three...