Building orientation is important in selective laser melting (SLM) processes. Current studies only focus on the horizontal and vertical building orientations without considering different modes of horizontal orientations. In fact, for horizontal orientation, different surfaces of the sample that contact the substrate will affect the heat transfer mode and efficiency, and in turn affect the microstructure and material properties. In this paper, the effect of two modes of horizontal building orientations on microstructure, mechanical and surface properties of SLM Ti6Al4V was studied. Current research about building orientation is deficient because the geometry of samples or test surfaces are not strictly defined, which seriously influences the results due to their different heat transfer efficiency and mode. Therefore, the geometry of the samples and test surfaces were clearly defined, and its necessity was proved in this study. To achieve the research goal, three test samples were prepared: sample SLM-PB-S with the building orientation parallel to the substrate and the shorter side L1 contacts it, sample SLM-PB-L with the building orientation parallel to the substrate and the longer side L2 contacts it and sample SLM-VB with the building orientation vertical to the substrate. Subsequently, the microstructure, grain information, densification, residual stress, micro-hardness, tensile properties and surface topography of different samples were analyzed and compared. In the results, SLM-PB-S exhibited denser microstructure and better mechanical properties than SLM-PB-L, including smaller grain size, stronger texture, higher density, micro-hardness, tensile strength, plasticity and better surface quality. It originates from a higher cooling rate and shorter scanning time between layers during SLM-PB-S fabrication, leading to finer grains, lower porosity and better interlayer metallurgical bonding, thus resulting in better material properties. This study can provide a reference to select the proper building orientation in SLM.