Abstract Two and three dimensional growth of SrTiO3 films on (001) MgO substrate was achieved by pulsed laser interval and pulsed laser deposition respectively. The growth mode was monitored by in-situ reflection high energy electron diffraction. Interval deposition forces layer-by-layer growth of materials even with such a large lattice misfit (~7.9%). A titanium dioxide buffer monolayer was deposited to allow the film to wet the substrate to encourage two dimensional growth of the strontium titanate. A variety of defects was investigated using transmission electron microscopy and high resolution scanning transmission electron microscopy. Misfit dislocations, steps at the interface, Ti-rich defects and regularly shaped nano-holes connected by anti-phase boundaries were found to be the dominant defects in these films grown layer by layer. The edges of the nano-holes were mainly along  and  for a  growth direction. The large strain between the two crystal systems with large lattice mismatch leads to in-plane tensile stress during the layer-by-layer growth. The stress is relieved in part by the holes. The films with a three dimensional growth mode possess a uniform surface with dislocations as the dominant defects. The individual densities of the various defects, including a Ti-rich phase and misfit and threading dislocations, are determined by the kinetics of the deposition method.