The University of Rochester was founded in 1850 as a Baptist-sponsored institution. The impetus to form the university came primarily from the town of Hamilton, New York, which has been home to Colgate University since 1819. In 1848, the Baptist Education Society planned to move Colgate University (then known as Madison University) to the city of Rochester, but was halted by legal action in Hamilton. Dissenting Colgate trustees, faculty, and students founded the University of Rochester with a charter granted from the Regents of the University of the State of New York on January 31, 1850. Classes began that November, with approximately 60 students enrolling, including 28 transfers from Madison.
The University of Rochester's campus was originally in downtown Rochester at the United States Hotel, which was located on Buffalo Street near Elizabeth Street, which today is West Main Street near the I-490 overpass. In 1853, the campus moved east to a then-suburban location on what is now University Avenue. Local businessman and Congressman Azariah Boody donated 8 acres (3.2 ha) of land for the new campus, and the University purchased a further 17 acres (6.9 ha) from him. UR would remain on this campus until the current River Campus was constructed in 1930, and the university continues to own a small part of the University Avenue campus (where the university-owned Memorial Art Gallery is located).
The first female students were admitted in 1900, the result of an effort led by Susan B. Anthony and Helen Barrett Montgomery. During the 1890s, a number of women took classes and labs at the university as "visitors" but were not officially enrolled nor were their records included in the college register. President David Jayne Hill allowed the first woman, Helen E. Wilkinson, to enroll as a normal student, although she was not allowed to matriculate or to pursue a degree. Thirty-three women enrolled among the first class in 1900, and Ella S. Wilcoxen was the first to receive a degree, in 1901. When the River Campus was completed in 1930, male students moved there while the female students remained on the University Avenue campus until 1955.