In May 1995 an impressive karstic cave possessing dozens of burials dating to the main phase of the Chalcolithic Period (ca. 4500–3500 bce) was discovered in Peqi'in in the high hills of Galilee in northern Israel. It was subsequently excavated over the course of the following months. The large amount of unique ceramic ossuaries and the variety of burial offerings shed light, for the first time, on this critical societal development period in a region where its remains have rarely been found. Although the major significance of the cave is as a mortuary center, it was also utilized in a domestic capacity some centuries previously, during the generally less familiar Early Chalcolithic Period. We present here the dating of 22 14C samples collected from these separate phases of cave use. Their interpretation should aid in understanding of the absolute dating of the Chalcolithic Period and its relation to the period preceding its inception.