Music had always played a major part in the society and culture of the Republic of Venice, and the 16th century was a time of great musical innovation. The Cathedral of San Marco was the center of musical activity, and since the Republic and the Cathedral came under the jurisdiction of the Doge--the chief magistrate--rather than any ecclesiastical authority, Venice developed very different musical and liturgical traditions from Rome. There were many parades and processions of state, fusing civic and religious elements, in which music performance was an integral part. The Scuole, confraternities which served as religious and charitable organizations, also played a vital role in the cultural and ceremonial life of the city. The largest and most important of these confraternities, the Scuole Grandi, maintained musical establishments for long periods of time, and employed musicians for their concerts, religious processions, and participation in state celebrations. Wealthy private citizens supported musicians, too, and many music-loving nobles became patrons for ensembles of musicians in the same way as the ruling houses of Mantua, Ferrara, or Urbino. A great number of people possessed musical instruments, and all the classes enjoyed making music and listening to it.

By the 16th century, Venice had also become an important center of music publishing, and composers came from all over Europe to benefit from the new technology of the printing press. Many printers were also active musicians and composers, and they played a direct role in the arranging of the pieces they published. In addition to being a center of musical excellence, Venice had many workshops for the manufacture and export of musical instruments. Still another event in the history of music in the 16th century was the growth of the Commedia dell'Arte, a form of improvisational theatre. After about 1550, troupes of traveling players began to perform in Lombardy and the Venetian Republic, and soon the best troupes were invited to play in courts all over Italy and Europe.

But it was the polyphonic style which developed at the Cathedral of San Marco which became Venice's greatest legacy to the history of music. Before the 9th century, plainsong, a type of chant consisting of a single, unaccompanied melodic line, sung in a free, rather than a measured rhythm, had been the liturgical music of the Roman Catholic Church. Then an early form of polyphony called organum developed, in which one voice at an interval was added to enhance the harmony. Over time, composers began to write more parts that were not just simple transpositions and created true polyphony, with several interweaving melodic lines. Between 1317 and 1318, Marchettus of Padova, a musical theorist and composer, wrote two major treatises on plainchant and polyphony. These treatises, which were widely copied and disseminated, proved to be greatly influential during the 14th and 15th centuries.

Polyphony became firmly established in Venice by way of the Franco-Flemish school. By the 15th century, musicians in the low countries (modern day Netherlands and Belgium) were already using polyphony in sacred music, and many came to the major centers of Italy: Rome, Ferrara, Florence, and Milan, where they influenced Italian composers. The election of Flemish composer Adrian Willaert as maestro di cappella of the Cathedral of San Marco in 1527 (a post he retained until his death in 1562) proved to be a turning point in the history of music. Employing San Marco's unique, spacious interior with opposing choir lofts, Willaert divided the choral body into two sections and wrote antiphonal music in which these choirs sang successive, often contrasting phrases of music, united by the sound of the organ. The effect proved to be magnificent, as the sound bounced off the main dome and choir area, aided by the capacious acoustical space. There may also have been additional platforms, or palchetti, built to accommodate instrumentalists during larger festivals. These 'cori spezzati,' or split choirs, represented a significant shift from former polyphonic compositions, and through his work as a musician, teacher, and composer, Willaert helped to establish the Venetian School--a group of outstanding composers including Cipriano de Rore, Giseffo Zarlino, Claudio Merlo, and Andrea and Giovanni Gabrieli. They created great works suited to Venice's unique cultural and religious traditions and received acclaim from all over Europe.