The earliest settlers in Mardin were Assyrians/Syriacs, arriving in the 3rd century AD; the old Assyro-Babylonian religion existed in Mardin until the 18th century. Most Syriac Orthodox churches and monasteries in the city, which are still active today, date from the 5th century AD, such as the Deyrülzafarân Monastery. Mardin is likely the Maride and Marida of the Greeks and Romans. Another important church, Kırklar Kilisesi (Church of the 40 Martyrs), originally built in the name of Benham and Saro, the two sons of the Assyrian ruler who executed them because they chose to become Christian, dates from 569 AD. , and even during its use as a capital by the Artukid Turkish dynasty which ruled Eastern Anatolia and Northern Mesopotamia between the twelfth and fourteenth centuries. The 12th century Sitti Radviyye Madrasa, the oldest of its kind in Anatolia, dates from this period.

The lands of the Artukid dynasty fell to the Mongols who took control of the region in 1394, but the Mongols never directly governed the area. Mardin was later controlled by the Turkish Akkoyunlu kingdom. The Kasımiye Madrasa was built by Sultan Kasım, son of the Akkoyunlu Sultan Cihangir, between 1457 and 1502.