He was born at Celleno and became the a pupil of a baptized Jew, Giovanni Battista, who instructed him in Hebrew. On completing his studies, Bartolocci entered the Cistercian order. It was from Battista that Bartolocci obtained his knowledge of Hebrew and rabbinical literature.
He was appointed, in 1651, professor of Hebrew and Rabbinics at the Collegium Neophytorum at Rome, and "Scriptor Hebraicus" at the Vatican Library. It was in the Vatican, and with the assistance of Battista, that Bartolocci received his preparation for the work that was to give him lasting fame in the world of Jewish bibliography; and it was at the Vatican and its subsidiary libraries that he obtained his chief materials.
In 1675 he began in Rome the publication of Bibliotheca Magna Rabbinica, a bibliography, in Latin and Hebrew, of Hebrew literature, arranged according to the names of the authors. This work appeared in four folio volumes, 1675-1693, three of which were published by the author and the fourth by Carlo Giuseppe Imbonati, his disciple. Imbonati's supplement contained a list of authors arranged according to the subjects on which they wrote. The latter added to this work a fifth volume, the Bibliotheca Latina Hebraica, Rome, 1694, which contained the works and the names of Christian authors who had written in Latin on Jews and Judaism.
It was from Battista that Bartolocci obtained the idea and plan of the Bibliotheca Magna Rabbinica, as well as part of the material. Battista began the composition of the book in a chronological order, which order was abandoned by Bartolocci. Richard Simon, in writing in his Bibliothèque Critique about Bartolocci's work, says:
Complaints were also made that he devoted space to refutations of Jewish arguments and that his translations from the Talmud were faulty. On the other hand, Johann Christoph Wolf attributes to Bartolocci the motive and stimulus for his own later work.