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The idea of an archaeological museum takes shape in Siena between the late 19th century and the early 20th century, within those institutions that, due to various circumstances, had seen the formation of small nuclei of antiquities.

The idea of an archaeological museum began to take shape in Siena between the late 19th and early 20th centuries, within those institutions that, due to various circumstances, had seen the formation of small collections of antiquities. The scientific planning of the museum was carried out in the 1920s by the young Ranuccio Bianchi Bandinelli. The museum began to take form in the premises on Via della Sapienza, where between 1931 and 1933, materials from the Accademia dei Fisiocritici (findings from the city and the Mieli collection, donated to the Municipality of Siena in 1882) and the Bargagli di Sarteano collection, transferred to Siena in August 1931, were displayed. The establishment of the Royal Archaeological Museum dates back to October 7, 1941. The archaeological collection continued to grow in the following years: in 1951, the collection of Bonaventura Chigi Zondadari was donated to the state; two years later, the state acquired the collection of Emilio Bonci Casuccini, which provided valuable documentation of the archaeology of Chiusi. In the second half of the 20th century, the museum expanded thanks to accidental discoveries from the surrounding area and excavations carried out by the Superintendence. In the late 1980s, the museum was transferred to the Santa Maria della Scala complex, occupying the spaces at the corner of the hospital, the ancient women's pilgrim area, and some adjacent rooms. In 2001, the museum was moved once more to the lowest levels of the Santa Maria (I-II), consisting of tunnels carved into sandstone and large brick-built spaces.

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