In a method of fabricating a semiconductor device having an alignment key and a semiconductor device fabricated thereby. The method of fabricating a semiconductor device includes providing a semiconductor substrate having a scribe lane region and a cell region. An etch barrier pattern and a gate pattern are formed on the scribe lane region and the cell region respectively. A first interlayer insulating layer is formed to cover the etch barrier pattern and the gate pattern. A preliminary alignment key pattern and a bit line pattern are formed on the first interlayer insulating layer of the scribe lane region and the cell region respectively. A second interlayer insulating layer is formed to cover the preliminary alignment key pattern and the bit line pattern. The second interlayer insulating layer and the first interlayer insulating layer are patterned to expose the etch barrier pattern, thereby forming an alignment key pattern in the scribe lane region, and concurrently, forming a storage node contact opening in the cell region.