We collected charcoal fragments during an archaeological excavation at the Tell Ghanem al-Ali site, located on the lowest terrace of the middle Euphrates River, and measured their radiocarbon ages with accelerator mass spectrometry (AMS). Two trenches, Square-1 and Square-2, were dug on the slope of the tell; 8 building levels were detected in the Square-2 trench. In total, 31 charcoal samples were collected from the 2 trenches, and their calibrated ages ranged from 3100–2900 cal BC at the lowest building level to 2400–2050 cal BC at the uppermost layers of the mound, and concentrated in the period 2650–2450 cal BC. The pottery fragments collected on the surface of the mound before the excavation survey was started, as well as those collected from the sediment layers during the excavation, were assigned on the basis of typological sequences to the Early Bronze Age (EB)-III and EB-IV periods. Thus, the concentrated dates (2650–2450 cal BC) obtained by 14C dating are consistent with the age estimated by archaeological contexts. However, the oldest dates of the lowest level (level-7) go back to 3100–2900 cal BC, and these dates may suggest the existence of the human residence prior to the EB period at the site, and may therefore lead to a revision of the oldest age limit of the EB period currently accepted in the region.