The size and shape Evolution of self-assembled InAs quantum dots (QDs) influenced by 2.0-ML InAs seed layer has been systematically investigated for 2.0, 2.5, and 2.9-ML deposition on GaAs(1 0 0) substrate. Based on comparisons with the evolution of InAs islands on single layer samples at late growth stage, the bimodal size distribution of InAs islands at 2.5-ML InAs coverage and the formation of larger InAs quantum dots at 2.9-ML deposition have been observed on the second InAs layer. The further cross-sectional transmission electron microscopy measurement indicates the larger InAs QDs: at 2.9-ML deposition on the second layer are free of dislocation. In addition, the interpretations for the size and shape evolution of InAs/GaAs QDs on the second layer will be presented.