A quantum dot light-emitting diode substrate having a bonding layer and a method of preparing the same are provided. The quantum dot light-emitting diode substrate including a plurality of sub-pixel light-emitting regions, wherein each of the sub-pixel light-emitting regions includes a light-emitting layer including a bonding layer and a quantum dot bonded to the bonding layer. The quantum dot light-emitting diode substrate can be prepared with high resolution by a convenient process, and is suitable for mass production.